α-synuclein核输入机制及功能的研究
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摘要
帕金森氏病(PD, parkinson's disease)是临床最常见的神经系统退行性疾病之一,65岁以上人群的发病率为1%,我国大约有170万帕金森氏病患者,随着人口的老龄化的加剧,发病人数还将呈现出逐年上升的趋势。到目前为止,在大多数情况下帕金森氏病的病因仍然不是很清楚。1913年Frederick Lewy首次证明路易氏小体(LB, Lewy body)是帕金森氏病病人所具备的病理特征。路易氏小体的主要成分是一种以非折叠形式存在并且具有高度聚集倾向性的蛋白,即a-synuclein。有很多研究表明,a-synuclein存在于神经元的胞核内。尽管如此,直至今日仍然没有a-synuclein在细胞核内确切功能的报道。是否核定位是a-synuclein的一种普遍特性,还是PD的病理结果,到目前为止仍然是个迷。
在本研究中,我们试图解决以下三个方面的问题：①对a-synuclein核输入起决定性作用的序列；②α-synuclein进入细胞核的方式；③a-synuclein在细胞核内对细胞功能的影响。
首先,我们应用现代生物信息学的方法对a-synuclein的核定位信号以及核输入相关的片段进行预测,并应用PCR (polymerase chain reaction)的方法获得一系列与a-synuclein核输入相关的片段,并构建入pEGFP-N1真核表达载体系统,转染PC12细胞后在细胞水平寻找对a-synuclein核输入起决定性作用的序列片段。
其次,在确定核输入方式上,我们采用了核输入抑制与RNA干扰(RNAi, RNA interference)相结合的方法,同时应用免疫荧光、免疫沉淀、实时荧光定量PCR (Real Time PCR, real-time quantitative polymerase chain reaction)以及WB (western blot)对a-synuclein的核输入方式进行验证。
最后,为了探讨a-synuclein在细胞核中所发挥的作用,我们分别构建了a-synuclein与一个核定位序列(NLS, nuclear localization signal)以及a-synuclein与一个核输出序列(NES, nuclear export signal)融合表达的载体。转染PC12细胞并经G418稳定筛选后获得了a-synuclein分别在细胞核或细胞浆集中分布的稳定细胞株,分别从生长特性、形态学分析、细胞周期分布、对神经毒性剂敏感性等方面研究a-synuclein在细胞核集中分布所发挥的作用。同时,我们利用酵母表达纯化了a-synuclein-GFP融合蛋白,并应用圆二色谱研究a-synuclein对DNA的影响。
结果表明,a-synuclein的N端和C端都在其核输入过程中发挥一定作用,并且其进入细胞核需要importin a的介导并在Ran水解GTP提供能量的情况下才能完成。a-synuclein进入细胞核后对细胞形态几乎没有影响,但会导致细胞生长速度减慢以及细胞周期的延迟,并进一步增加细胞对神经毒性剂MPP+(1-methyl-4-phenylpyridinium)的敏感性。另外,圆二色谱结果表明,两个α-synuclein分子能够与一个类GC box样单核苷酸序列结合。
综上所述,α-synuclein可能在其N端和C端片段共同介导下,首先与importinα结合,并进一步与importinβ结合后形成核输入复合物,在Ran水解GTP提供能量的情况下,通过核孔复合物进入到细胞核内。α-synuclein进入细胞核后,能与转录因子SP1 (specificity protein 1)竞争性结合启动子区上富含GC的区域,从而导致转录因子SPl所介导的E2F家族蛋白与启动子区上富含GC的DNA序列相结合并进一步形成cyclins和cdks复合物的过程受阻,从而引起细胞周期调控的异常,进一步导致其对神经毒性剂MPP+的敏感性增加。
PD (parkinson's disease) is one of the most common neurodegenerative diseases, which affect 1%of the people over 65 years old. There are 170 million PD patients in our country, the figure will increase year by year with the aggravating of aging. So far, the pathogen of PD is unclear in most case. The LB, which was first described by Frederick Lewy in 1913, is present in essentially all cases of PD. The major protein constituent of LBs is alpha-synuclein (a-synuclein), a natively unfolded protein having high propensity for aggregation. Several studies have shown the presence of a-synuclein in the neuronal nuclei. However, no definite nuclear functions have been attributed to a-synuclein to date. It is not known if nuclear localization is the common property of a-synuclein or it is cause/consequence of PD pathology.
In this study, we sought to address the following three aspects:the protein domains responsible for the nuclear import of a-synuclein; the way of a-synuclein import into the nuclear; the effect on cell function of a-synuclein in the nuclear.
First of all, the bioinformatics were used to predict both the NLS in a-synuclein and the protein domains responsible for the nuclear localization of a-synuclein. Using deletion mutant forms of a-synuclein, we identified protein domains that are responsible for the observed nuclear import of a-synuclein in PC 12 cells.
Secondly, the nuclear import inhibition、RNAi (RNA interference)-. Immunofluorescence、Immunoprecipitatio、Real Time PCR (real-time quantitative polymerase chain reaction) and WB (western blot), were used to explore the way of a-synuclein import into the nuclei.
Finally, in order to examine the ability of a-synuclein in the nucleus, we created constructs of a-synuclein tagged with either a nuclear localization sequence (NLS) or a nuclear export sequence (NES). We used these constructs to transfect PC12 cells, and screened with G418, then the stable cell lines which express a-synuclein are located in the nucleus or cytoplasm were acquired. The growth curve morphological analysis、cell cycle sensitivity to neurotoxic agents (MPP+) of the stable cell lines were analyzed. The a-synuclein-GFP fusion protein was expressed and purified with yeast, then the circular dichroism (CD) was used to study the effect of a-synuclein on DNA.
The results showed that, both the N terminal and the C terminal were responsible for the nuclear import of a-synuclein. a-synuclein could import into the nuclei through nuclear pore complex (NPC), mediated by importin a and provided energy by hydrolysis of GTP. There was no effect on cell morphological whenα-synuclein import into the nuclei, but both the growth curve and the cell cycle were delayed. And the PC 12 cells with a-synuclein localization in nuclei were sensitivity to neurotoxic agents (MPP+). Moreover, the results of the circular dichroism showed that twoα-synuclein molecules were prone to combine with one GC box-like single nucleotide sequence.
In summary, mediated by both the N terminal and the C terminal,α-synuclein could combine with importin a and importinβ, then import into the nuclei through nuclear pore complex (NPC), provided energy by hydrolysis of GTP.α-synuclein could compete with the transcription factor SP1 for the same GC rich region in promoter, then the process of both the E2F family protein interaction with the GC rich region mediated by the transcription factor SP1 and the cyclins/cdks complex formation were inhibited, so the cell cycle was disturbed, it led to the PC 12 cells with a-synuclein localization in nuclei more sensitivity to neurotoxic agents (MPP+).
在本研究中,我们试图解决以下三个方面的问题：①对a-synuclein核输入起决定性作用的序列；②α-synuclein进入细胞核的方式；③a-synuclein在细胞核内对细胞功能的影响。
首先,我们应用现代生物信息学的方法对a-synuclein的核定位信号以及核输入相关的片段进行预测,并应用PCR (polymerase chain reaction)的方法获得一系列与a-synuclein核输入相关的片段,并构建入pEGFP-N1真核表达载体系统,转染PC12细胞后在细胞水平寻找对a-synuclein核输入起决定性作用的序列片段。
其次,在确定核输入方式上,我们采用了核输入抑制与RNA干扰(RNAi, RNA interference)相结合的方法,同时应用免疫荧光、免疫沉淀、实时荧光定量PCR (Real Time PCR, real-time quantitative polymerase chain reaction)以及WB (western blot)对a-synuclein的核输入方式进行验证。
最后,为了探讨a-synuclein在细胞核中所发挥的作用,我们分别构建了a-synuclein与一个核定位序列(NLS, nuclear localization signal)以及a-synuclein与一个核输出序列(NES, nuclear export signal)融合表达的载体。转染PC12细胞并经G418稳定筛选后获得了a-synuclein分别在细胞核或细胞浆集中分布的稳定细胞株,分别从生长特性、形态学分析、细胞周期分布、对神经毒性剂敏感性等方面研究a-synuclein在细胞核集中分布所发挥的作用。同时,我们利用酵母表达纯化了a-synuclein-GFP融合蛋白,并应用圆二色谱研究a-synuclein对DNA的影响。
结果表明,a-synuclein的N端和C端都在其核输入过程中发挥一定作用,并且其进入细胞核需要importin a的介导并在Ran水解GTP提供能量的情况下才能完成。a-synuclein进入细胞核后对细胞形态几乎没有影响,但会导致细胞生长速度减慢以及细胞周期的延迟,并进一步增加细胞对神经毒性剂MPP+(1-methyl-4-phenylpyridinium)的敏感性。另外,圆二色谱结果表明,两个α-synuclein分子能够与一个类GC box样单核苷酸序列结合。
综上所述,α-synuclein可能在其N端和C端片段共同介导下,首先与importinα结合,并进一步与importinβ结合后形成核输入复合物,在Ran水解GTP提供能量的情况下,通过核孔复合物进入到细胞核内。α-synuclein进入细胞核后,能与转录因子SP1 (specificity protein 1)竞争性结合启动子区上富含GC的区域,从而导致转录因子SPl所介导的E2F家族蛋白与启动子区上富含GC的DNA序列相结合并进一步形成cyclins和cdks复合物的过程受阻,从而引起细胞周期调控的异常,进一步导致其对神经毒性剂MPP+的敏感性增加。
PD (parkinson's disease) is one of the most common neurodegenerative diseases, which affect 1%of the people over 65 years old. There are 170 million PD patients in our country, the figure will increase year by year with the aggravating of aging. So far, the pathogen of PD is unclear in most case. The LB, which was first described by Frederick Lewy in 1913, is present in essentially all cases of PD. The major protein constituent of LBs is alpha-synuclein (a-synuclein), a natively unfolded protein having high propensity for aggregation. Several studies have shown the presence of a-synuclein in the neuronal nuclei. However, no definite nuclear functions have been attributed to a-synuclein to date. It is not known if nuclear localization is the common property of a-synuclein or it is cause/consequence of PD pathology.
In this study, we sought to address the following three aspects:the protein domains responsible for the nuclear import of a-synuclein; the way of a-synuclein import into the nuclear; the effect on cell function of a-synuclein in the nuclear.
First of all, the bioinformatics were used to predict both the NLS in a-synuclein and the protein domains responsible for the nuclear localization of a-synuclein. Using deletion mutant forms of a-synuclein, we identified protein domains that are responsible for the observed nuclear import of a-synuclein in PC 12 cells.
Secondly, the nuclear import inhibition、RNAi (RNA interference)-. Immunofluorescence、Immunoprecipitatio、Real Time PCR (real-time quantitative polymerase chain reaction) and WB (western blot), were used to explore the way of a-synuclein import into the nuclei.
Finally, in order to examine the ability of a-synuclein in the nucleus, we created constructs of a-synuclein tagged with either a nuclear localization sequence (NLS) or a nuclear export sequence (NES). We used these constructs to transfect PC12 cells, and screened with G418, then the stable cell lines which express a-synuclein are located in the nucleus or cytoplasm were acquired. The growth curve morphological analysis、cell cycle sensitivity to neurotoxic agents (MPP+) of the stable cell lines were analyzed. The a-synuclein-GFP fusion protein was expressed and purified with yeast, then the circular dichroism (CD) was used to study the effect of a-synuclein on DNA.
The results showed that, both the N terminal and the C terminal were responsible for the nuclear import of a-synuclein. a-synuclein could import into the nuclei through nuclear pore complex (NPC), mediated by importin a and provided energy by hydrolysis of GTP. There was no effect on cell morphological whenα-synuclein import into the nuclei, but both the growth curve and the cell cycle were delayed. And the PC 12 cells with a-synuclein localization in nuclei were sensitivity to neurotoxic agents (MPP+). Moreover, the results of the circular dichroism showed that twoα-synuclein molecules were prone to combine with one GC box-like single nucleotide sequence.
In summary, mediated by both the N terminal and the C terminal,α-synuclein could combine with importin a and importinβ, then import into the nuclei through nuclear pore complex (NPC), provided energy by hydrolysis of GTP.α-synuclein could compete with the transcription factor SP1 for the same GC rich region in promoter, then the process of both the E2F family protein interaction with the GC rich region mediated by the transcription factor SP1 and the cyclins/cdks complex formation were inhibited, so the cell cycle was disturbed, it led to the PC 12 cells with a-synuclein localization in nuclei more sensitivity to neurotoxic agents (MPP+).
引文
[1]Parkinson J. An essay on the shaking palsy.1817. J Neuropsychiatry Clin Neurosci 2002;14(2):223-36; discussion 22.
[2]Gasser T. Genetics of Parkinson's disease. J Neurol 2001;248(10):833-40.
[3]Maroteaux L, Campanelli JT, Scheller RH. Synuclein:a neuron-specific protein localized to the nucleus and presynaptic nerve terminal. J Neurosci 1988;8(8):2804-15.
[4]Conway KA, Harper JD, Lansbury PT, Jr. Fibrils formed in vitro from alpha-synuclein and two mutant forms linked to Parkinson's disease are typical amyloid. Biochemistry 2000;39(10):2552-63.
[5]El-Agnaf OM, Jakes R, Curran MD, Wallace A. Effects of the mutations Ala30 to Pro and Ala53 to Thr on the physical and morphological properties of alpha-synuclein protein implicated in Parkinson's disease. FEBS Lett 1998;440(1-2):67-70.
[6]Bisaglia M, Schievano E, Caporale A, Peggion E, Mammi S. The 11-mer repeats of human alpha-synuclein in vesicle interactions and lipid composition discrimination:a cooperative role. Biopolymers 2006;84(3):310-6.
[7]Davidson WS, Jonas A, Clayton DF, George JM. Stabilization of alpha-synuclein secondary structure upon binding to synthetic membranes. J Biol Chem 1998;273(16):9443-9.
[8]Jao CC, Der-Sarkissian A, Chen J, Langen R. Structure of membrane-bound alpha-synuclein studied by site-directed spin labeling. Proc Natl Acad Sci U S A 2004; 101(22):8331-6.
[9]Biere AL, Wood SJ, Wypych J, Steavenson S, Jiang Y, Anafi D, et al. Parkinson's disease-associated alpha-synuclein is more fibrillogenic than beta-and gamma-synuclein and cannot cross-seed its homologs. J Biol Chem 2000;275(44):34574-9.
[10]Perrin RJ, Woods WS, Clayton DF, George JM. Interaction of human alpha-Synuclein and Parkinson's disease variants with phospholipids. Structural analysis using site-directed mutagenesis. J Biol Chem 2000;275(44):34393-8.
[11]Weinreb PH, Zhen W, Poon AW, Conway KA, Lansbury PT, Jr. NACP, a protein implicated in Alzheimer's disease and learning, is natively unfolded. Biochemistry 1996;35(43):13709-15.
[12]Bussell R, Jr., Eliezer D. Effects of Parkinson's disease-linked mutations on the structure of lipid-associated alpha-synuclein. Biochemistry 2004;43(16):4810-8.
[13]Chandra S, Chen X, Rizo J, Jahn R, Sudhof TC. A broken alpha-helix in folded alpha-Synuclein. J Biol Chem 2003;278(17):15313-8.
[14]Trojanowski JQ, Lee VM. Parkinson's disease and related alpha-synucleinopathies are brain amyloidoses. Ann N Y Acad Sci 2003;991:107-10.
[15]Barghorn S, Davies P, Mandelkow E. Tau paired helical filaments from Alzheimer's disease brain and assembled in vitro are based on beta-structure in the core domain. Biochemistry 2004;43(6):1694-703.
[16]Giasson BI, Uryu K, Trojanowski JQ, Lee VM. Mutant and wild type human alpha-synucleins assemble into elongated filaments with distinct morphologies in vitro. J Biol Chem 1999;274(12):7619-22.
[17]Serpell LC, Berriman J, Jakes R, Goedert M, Crowther RA. Fiber diffraction of synthetic alpha-synuclein filaments shows amyloid-like cross-beta conformation. Proc Natl Acad Sci U S A 2000;97(9):4897-902.
[18]Kruger R, Kuhn W, Muller T, Woitalla D, Graeber M, Kosel S, et al. Ala30Pro mutation in the gene encoding alpha-synuclein in Parkinson's disease. Nat Genet 1998; 18(2):106-8.
[19]Polymeropoulos MH, Lavedan C, Leroy E, Ide SE, Dehejia A, Dutra A, et al. Mutation in the alpha-synuclein gene identified in families with Parkinson's disease. Science 1997;276(5321):2045-7.
[20]Zarranz JJ, Alegre J, Gomez-Esteban JC, Lezcano E, Ros R, Ampuero I, et al. The new mutation, E46K, of alpha-synuclein causes Parkinson and Lewy body dementia. Ann Neurol 2004;55(2):164-73.
[21]Greenbaum EA, Graves CL, Mishizen-Eberz AJ, Lupoli MA, Lynch DR, Englander SW, et al. The E46K mutation in alpha-synuclein increases amyloid fibril formation. J Biol Chem 2005;280(9):7800-7.
[22]Perrin RJ, Woods WS, Clayton DF, George JM. Exposure to long chain polyunsaturated fatty acids triggers rapid multimerization of synucleins. J Biol Chem 2001;276(45):41958-62.
[23]Li J, Uversky VN, Fink AL. Effect of familial Parkinson's disease point mutations A30P and A53T on the structural properties, aggregation, and fibrillation of human alpha-synuclein. Biochemistry 2001;40(38):11604-13.
[24]Smith WW, Margolis RL, Li X, Troncoso JC, Lee MK, Dawson VL, et al. Alpha-synuclein phosphorylation enhances eosinophilic cytoplasmic inclusion formation in SH-SY5Y cells. J Neurosci 2005;25(23):5544-52.
[25]Unger EL, Eve DJ, Perez XA, Reichenbach DK, Xu Y, Lee MK, et al. Locomotor hyperactivity and alterations in dopamine neurotransmission are associated with overexpression of A53T mutant human alpha-synuclein in mice. Neurobiol Dis 2006;21(2):431-43.
[26]Pandey N, Schmidt RE, Galvin JE. The alpha-synuclein mutation E46K promotes aggregation in cultured cells. Exp Neurol 2006; 197(2):515-20.
[27]Chen L, Feany MB. Alpha-synuclein phosphorylation controls neurotoxicity and inclusion formation in a Drosophila model of Parkinson disease. Nat Neurosci 2005;8(5):657-63.
[28]Choi W, Zibaee S, Jakes R, Serpell LC, Davletov B, Crowther RA, et al. Mutation E46K increases phospholipid binding and assembly into filaments of human alpha-synuclein. FEBS Lett 2004;576(3):363-8.
[29]Conway KA, Rochet JC, Bieganski RM, Lansbury PT, Jr. Kinetic stabilization of the alpha-synuclein protofibril by a dopamine-alpha-synuclein adduct. Science 2001;294(5545):1346-9.
[30]Lavedan C. The synuclein family. Genome Res 1998;8(9):871-80.
[31]Hsu LJ, Mallory M, Xia Y, Veinbergs I, Hashimoto M, Yoshimoto M, et al. Expression pattern of synucleins (non-Abeta component of Alzheimer's disease amyloid precursor protein/alpha-synuclein) during murine brain development. J Neurochem 1998;71(1):338-44.
[32]Bayer TA, Jakala P, Hartmann T, Egensperger R, Buslei R, Falkai P, et al. Neural expression profile of alpha-synuclein in developing human cortex. Neuroreport 1999;10(13):2799-803.
[33]Galvin JE, Schuck TM, Lee VM, Trojanowski JQ. Differential expression and distribution of alpha-, beta-, and gamma-synuclein in the developing human substantia nigra. Exp Neurol 2001;168(2):347-55.
[34]Petersen K, Olesen OF, Mikkelsen JD. Developmental expression of alpha-synuclein in rat hippocampus and cerebral cortex. Neuroscience 1999;91(2):651-9.
[35]Stefanis L, Kholodilov N, Rideout HJ, Burke RE, Greene LA. Synuclein-1 is selectively up-regulated in response to nerve growth factor treatment in PC 12 cells. J Neurochem 2001;76(4):1165-76.
[36]Vila M, Vukosavic S, Jackson-Lewis V, Neystat M, Jakowec M, Przedborski S. Alpha-synuclein up-regulation in substantia nigra dopaminergic neurons following administration of the parkinsonian toxin MPTP. J Neurochem 2000;74(2):721-9.
[37]Hashimoto M, Yoshimoto M, Sisk A, Hsu LJ, Sundsmo M, Kittel A, et al. NACP, a synaptic protein involved in Alzheimer's disease, is differentially regulated during megakaryocyte differentiation. Biochem Biophys Res Commun 1997;237(3):611-6.
[38]Cole NB, Murphy DD. The cell biology of alpha-synuclein:a sticky problem? Neuromolecular Med 2002; 1(2):95-109.
[39]Zhang L, Zhang C, Zhu Y, Cai Q, Chan P, Ueda K, et al. Semi-quantitative analysis of alpha-synuclein in subcellular pools of rat brain neurons:an immunogold electron microscopic study using a C-terminal specific monoclonal antibody. Brain Res 2008; 1244:40-52.
[40]Schneider BL, Seehus CR, Capowski EE, Aebischer P, Zhang SC, Svendsen CN. Over-expression of alpha-synuclein in human neural progenitors leads to specific changes in fate and differentiation. Hum Mol Genet 2007; 16(6):651-66.
[41]Andringa G, Du F, Chase TN, Bennett MC. Mapping of rat brain using the Synuclein-1 monoclonal antibody reveals somatodendritic expression of alpha-synuclein in populations of neurons homologous to those vulnerable to Lewy body formation in human synucleopathies. J Neuropathol Exp Neurol 2003;62(10):1060-75.
[42]Mori F, Tanji K, Yoshimoto M, Takahashi H, Wakabayashi K. Immunohistochemical comparison of alpha- and beta-synuclein in adult rat central nervous system. Brain Res 2002;941 (1-2):118-26.
[43]Specht CG, Tigaret CM, Rast GF, Thalhammer A, Rudhard Y, Schoepfer R. Subcellular localisation of recombinant alpha- and gamma-synuclein. Mol Cell Neurosci 2005;28(2):326-34.
[44]Takahashi M, Kanuka H, Fujiwara H, Koyama A, Hasegawa M, Miura M, et al. Phosphorylation of alpha-synuclein characteristic of synucleinopathy lesions is recapitulated in alpha-synuclein transgenic Drosophila. Neurosci Lett 2003;336(3):155-8.
[45]Masliah E, Rockenstein E, Veinbergs I, Mallory M, Hashimoto M, Takeda A, et al. Dopaminergic loss and inclusion body formation in alpha-synuclein mice: implications for neurodegenerative disorders. Science 2000;287(5456):1265-9.
[46]Goers J, Manning-Bog AB, McCormack AL, Millett IS, Doniach S, Di Monte DA, et al. Nuclear localization of alpha-synuclein and its interaction with histones. Biochemistry 2003;42(28):8465-71.
[47]McLean PJ, Ribich S, Hyman BT. Subcellular localization of alpha-synuclein in primary neuronal cultures:effect of missense mutations. J Neural Transm Suppl 2000(58):53-63.
[48]Yu S, Zuo X, Li Y, Zhang C, Zhou M, Zhang YA, et al. Inhibition of tyrosine hydroxylase expression in alpha-synuclein-transfected dopaminergic neuronal cells. Neurosci Lett 2004;367(1):34-9.
[49]Yu S, Li X, Liu G, Han J, Zhang C, Li Y, et al. Extensive nuclear localization of alpha-synuclein in normal rat brain neurons revealed by a novel monoclonal antibody. Neuroscience 2007;145(2):539-55.
[50]Leng Y, Chase TN, Bennett MC. Muscarinic receptor stimulation induces translocation of an alpha-synuclein oligomer from plasma membrane to a light vesicle fraction in cytoplasm. J Biol Chem 2001;276(30):28212-8.
[51]Hegde ML, Jagannatha Rao KS. Challenges and complexities of alpha-synuclein toxicity:new postulates in unfolding the mystery associated with Parkinson's disease. Arch Biochem Biophys 2003;418(2):169-78.
[52]Hegde ML, Rao KS. DNA induces folding in alpha-synuclein:understanding the mechanism using chaperone property of osmolytes. Arch Biochem Biophys 2007;464(1):57-69.
[53]Dev KK, Hofele K, Barbieri S, Buchman VL, van der Putten H. Part II: alpha-synuclein and its molecular pathophysiological role in neurodegenerative disease. Neuropharmacology 2003;45(1):14-44.
[54]Iwai A, Masliah E, Yoshimoto M, Ge N, Flanagan L, de Silva HA, et al. The precursor protein of non-A beta component of Alzheimer's disease amyloid is a presynaptic protein of the central nervous system. Neuron 1995;14(2):467-75.
[55]Spillantini MG, Schmidt ML, Lee VM, Trojanowski JQ, Jakes R, Goedert M. Alpha-synuclein in Lewy bodies. Nature 1997;388(6645):839-40.
[56]Wakabayashi K, Matsumoto K, Takayama K, Yoshimoto M, Takahashi H. NACP, a presynaptic protein, immunoreactivity in Lewy bodies in Parkinson's disease. Neurosci Lett 1997;239(1):45-8.
[57]Dingwall C, Sharnick SV, Laskey RA. A polypeptide domain that specifies migration of nucleoplasmin into the nucleus. Cell 1982;30(2):449-58.
[58]Kalderon D, Roberts BL, Richardson WD, Smith AE. A short amino acid sequence able to specify nuclear location. Cell 1984;39(3 Pt 2):499-509.
[59]Dingwall C, Laskey RA. Nuclear targeting sequences--a consensus? Trends Biochem Sci 1991;16(12):478-81.
[60]Dang CV, Lee WM. Identification of the human c-myc protein nuclear translocation signal. Mol Cell Biol 1988;8(10):4048-54.
[61]Zhou M, Liu H, Xu X, Zhou H, Li X, Peng C, et al. Identification of nuclear localization signal that governs nuclear import of BRD7 and its essential roles in inhibiting cell cycle progression. J Cell Biochem 2006;98(4):920-30.
[62]Hanaka H, Shimizu T, Izumi T. Nuclear-localization-signal-dependent and nuclear-export-signal-dependent mechanisms determine the localization of 5-lipoxygenase. Biochem J 2002;361(Pt 3):505-14.
[63]Antoine M, Reimers K, Wirz W, Gressner AM, Muller R, Kiefer P. Identification of an unconventional nuclear localization signal in human ribosomal protein S2. Biochem Biophys Res Commun 2005;335(l):146-53.
[64]Chan CK, Jans DA. Using nuclear targeting signals to enhance non-viral gene ransfer. Immunol Cell Biol 2002;80(2):119-30.
[65]Robbins J, Dilworth SM, Laskey RA, Dingwall C. Two interdependent basic domains in nucleoplasmin nuclear targeting sequence:identification of a class of bipartite nuclear targeting sequence. Cell 1991;64(3):615-23.
[66]Gasiorowski JZ, Dean DA. Mechanisms of nuclear transport and interventions. Adv Drug Deliv Rev 2003;55(6):703-16.
[67]Kotera I, Sekimoto T, Miyamoto Y, Saiwaki T, Nagoshi E, Sakagami H, et al. Importin alpha transports CaMKIV to the nucleus without utilizing importin beta. EmboJ2005;24(5):942-51.
[68]Jensen KF, Ohmstede CA, Fisher RS, Sahyoun N. Nuclear and axonal localization of Ca2+/calmodulin-dependent protein kinase type Gr in rat cerebellar cortex. Proc Natl Acad Sci U S A 1991;88(7):2850-3.
[69]Gorlich D, Kutay U. Transport between the cell nucleus and the cytoplasm. Annu Rev Cell Dev Biol 1999; 15:607-60.
[70]Komeili A, O'Shea EK. New perspectives on nuclear transport. Annu Rev Genet 2001;35:341-64.
[71]Nakielny S, Dreyfuss G. Import and export of the nuclear protein import receptor transportin by a mechanism independent of GTP hydrolysis. Curr Biol 1998;8(2):89-95.
[72]Ben-Ze'ev A, Shtutman M, Zhurinsky J. The integration of cell adhesion with gene expression:the role of beta-catenin. Exp Cell Res 2000;261(1):75-82.
[73]Fagotto F, Gluck U, Gumbiner BM. Nuclear localization signal-independent and importin/karyopherin-independent nuclear import of beta-catenin. Curr Biol 1998;8(4):181-90.
[74]Sachdev S, Bagchi S, Zhang DD, Mings AC, Hannink M. Nuclear import of IkappaBalpha is accomplished by a ran-independent transport pathway. Mol Cell Biol2000;20(5):1571-82.
[75]Sachdev S, Hoffmann A, Hannink M. Nuclear localization of IkappaB alpha is mediated by the second ankyrin repeat:the IkappaB alpha ankyrin repeats define a novel class of cis-acting nuclear import sequences. Mol Cell Biol 1998;18(5):2524-34.
[76]Hicks GR, Raikhel NV. Protein import into the nucleus:an integrated view. Annu Rev Cell Dev Biol 1995; 11:155-88.
[77]Kosugi S, Hasebe M, Entani T, Takayama S, Tomita M, Yanagawa H. Design of peptide inhibitors for the importin alpha/beta nuclear import pathway by activity-based profiling. Chem Biol 2008;15(9):940-9.
[78]Kontopoulos E, Parvin JD, Feany MB. Alpha-synuclein acts in the nucleus to inhibit histone acetylation and promote neurotoxicity. Hum Mol Genet 2006;15(20):3012-23.
[79]Baptista MJ, O'Farrell C, Daya S, Ahmad R, Miller DW, Hardy J, et al. Co-ordinate transcriptional regulation of dopamine synthesis genes by alpha-synuclein in human neuroblastoma cell lines. J Neurochem 2003;85(4):957-68.
[80]Lee MK, Stirling W, Xu Y, Xu X, Qui D, Mandir AS, et al. Human alpha-synuclein-harboring familial Parkinson's disease-linked Ala-53--> Thr mutation causes neurodegenerative disease with alpha-synuclein aggregation in transgenic mice. Proc Natl Acad Sci U S A 2002;99(13):8968-73.
[81]Cherny D, Hoyer W, Subramaniam V, Jovin TM. Double-stranded DNA stimulates the fibrillation of alpha-synuclein in vitro and is associated with the mature fibrils:an electron microscopy study. J Mol Biol 2004;344(4):929-38.
[82]Yamada M, Iwatsubo T, Mizuno Y, Mochizuki H. Overexpression of alpha-synuclein in rat substantia nigra results in loss of dopaminergic neurons, phosphorylation of alpha-synuclein and activation of caspase-9:resemblance to pathogenetic changes in Parkinson's disease. J Neurochem 2004;91 (2):451-61.
[83]Chesselet MF In vivo alpha-synuclein overexpression in rodents:a useful model of Parkinson's disease? ExpNeurol 2008;209(1):22-7.
[84]Lucking CB, Brice A. Alpha-synuclein and Parkinson's disease. Cell Mol Life Sci 2000;57(13-14):1894-908.
[85]Ostrerova N, Petrucelli L, Farrer M, Mehta N, Choi P, Hardy J, et al. alpha-Synuclein shares physical and functional homology with 14-3-3 proteins. J Neurosci 1999; 19(14):5782-91.
[86]Jenco JM, Rawlingson A, Daniels B, Morris AJ. Regulation of phospholipase D2: selective inhibition of mammalian phospholipase D isoenzymes by alpha-and beta-synucleins. Biochemistry 1998;37(14):4901-9.
[87]Klein J, Lindmar R, Loffelholz K. Muscarinic activation of phosphatidylcholine hydrolysis. Prog Brain Res 1996;109:201-8.
[88]Collas P, Husebye H, Alestrom P. The nuclear localization sequence of the SV40 T antigen promotes transgene uptake and expression in zebrafish embryo nuclei. Transgenic Res 1996;5(6):451-8.
[89]Xiao CY, Jans DA. An engineered site for protein kinase C flanking the SV40 large T-antigen NLS confers phorbol ester-inducible nuclear import. FEBS Lett 1998;436(3):313-7.
[90]Fukuda M, Gotoh I, Gotoh Y, Nishida E. Cytoplasmic localization of mitogen-activated protein kinase kinase directed by its NH2-terminal, leucine-rich short amino acid sequence, which acts as a nuclear export signal. J BiolChem 1996;271(33):20024-8.
[91]Nakamura S, Kawamoto Y, Nakano S, Akiguchi I, Kimura J. p35nck5a and cyclin-dependent kinase 5 colocalize in Lewy bodies of brains with Parkinson's disease. Acta Neuropathol 1997;94(2):153-7.
[92]Neystat M, Rzhetskaya M, Oo TF, Kholodilov N, Yarygina O, Wilson A, et al. Expression of cyclin-dependent kinase 5 and its activator p35 in models of induced apoptotic death in neurons of the substantia nigra in vivo. J Neurochem 2001;77(6):1611-25.
[93]Staropoli JF, McDermott C, Martinat C, Schulman B, Demireva E, Abeliovich A. Parkin is a component of an SCF-like ubiquitin ligase complex and protects postmitotic neurons from kainate excitotoxicity. Neuron 2003;37(5):735-49.
[94]Dauer W, Przedborski S. Parkinson's disease:mechanisms and models. Neuron 2003;39(6):889-909.
[95]Jordan-Sciutto KL, Dorsey R, Chalovich EM, Hammond RR, Achim CL. Expression patterns of retinoblastoma protein in Parkinson disease. J Neuropathol Exp Neurol 2003;62(1):68-74.
[96]El-Khodor BF, Oo TF, Kholodilov N, Burke RE. Ectopic expression of cell cycle markers in models of induced programmed cell death in dopamine neurons of the rat substantia nigra pars compacta. Exp Neurol 2003; 179(1):17-27.
[97]Hoglinger GU, Breunig JJ, Depboylu C, Rouaux C, Michel PP, Alvarez-Fischer D, et al. The pRb/E2F cell-cycle pathway mediates cell death in Parkinson's disease. Proc Natl Acad Sci U S A 2007;104(9):3585-90.
[98]Alvira D, Tajes M, Verdaguer E, de Arriba SG, Allgaier C, Matute C, et al. Inhibition of cyclin-dependent kinases is neuroprotective in l-methyl-4-phenylpyridinium-induced apoptosis in neurons. Neuroscience 2007;146(1):350-65.
[99]Lotharius J, O'Malley KL. The parkinsonism-inducing drug l-methyl-4-phenylpyridinium triggers intracellular dopamine oxidation. A novel mechanism of toxicity. J Biol Chem 2000;275(49):38581-8.
[100]Lee M, Hyun D, Halliwell B, Jenner P. Effect of the overexpression of wild-type or mutant alpha-synuclein on cell susceptibility to insult. J Neurochem 2001;76(4):998-1009.
[101]Wu F, Poon WS, Lu G, Wang A, Meng H, Feng L, et al. Alpha-synuclein knockdown attenuates MPP+induced mitochondrial dysfunction of SH-SY5Y cells. Brain Res 2009; 1292:173-9.
[102]Fountaine TM, Venda LL, Warrick N, Christian HC, Brundin P, Channon KM, et al. The effect of alpha-synuclein knockdown on MPP+toxicity in models of human neurons. Eur J Neurosci 2008;28(12):2459-73.
[103]Fountaine TM, Wade-Martins R. RNA interference-mediated knockdown of alpha-synuclein protects human dopaminergic neuroblastoma cells from MPP(+) toxicity and reduces dopamine transport. J Neurosci Res 2007;85(2):351-63.
[104]Pizarro JG, Junyent F, Verdaguer E, Jordan J, Beas-Zarate C, Pallas M, et al. Effects of MPP+on the molecular pathways involved in cell cycle control in B65 neuroblastoma cells. Pharmacol Res 2010;61(5):391-9.
[105]Bai J, Nakamura H, Ueda S, Kwon YW, Tanaka T, Ban S, et al. Proteasome-dependent degradation of cyclin D1 in 1-methyl-4-phenylpyridinium ion (MPP+)-induced cell cycle arrest. J Biol Chem 2004;279(37):38710-4.
[106]Gomez-Tortosa E, Newell K, Irizarry MC, Sanders JL, Hyman BT. alpha-Synuclein immunoreactivity in dementia with Lewy bodies: morphological staging and comparison with ubiquitin immunostaining. Acta Neuropathol 2000;99(4):352-7.
[107]Tanji K, Mori F, Imaizumi T, Yoshida H, Matsumiya T, Tamo W, et al. Upregulation of alpha-synuclein by lipopolysaccharide and interleukin-1 in human macrophages. Pathol Int 2002;52(9):572-7.
[108]Sangchot P, Sharma S, Chetsawang B, Porter J, Govitrapong P, Ebadi M. Deferoxamine attenuates iron-induced oxidative stress and prevents mitochondrial aggregation and alpha-synuclein translocation in SK-N-SH cells in culture. Dev Neurosci 2002;24(2-3):143-53.
[109]Lin WL, DeLucia MW, Dickson DW. Alpha-synuclein immunoreactivity in neuronal nuclear inclusions and neurites in multiple system atrophy. Neurosci Lett 2004;354(2):99-102.
[110]Hegde ML, Anitha S, Latha KS, Mustak MS, Stein R, Ravid R, et al. First evidence for helical transitions in supercoiled DNA by amyloid Beta Peptide (1-42) and aluminum:a new insight in understanding Alzheimer's disease. J Mol Neurosci 2004;22(1-2):19-31.
[111]Suram A, Hegde ML, Rao KS. A new evidence for DNA nicking property of amyloid beta-peptide (1-42):relevance to Alzheimer's disease. Arch Biochem Biophys 2007;463(2):245-52.
[112]Roehl HH, Conrad SE. Identification of a G1-S-phase-regulated region in the human thymidine kinase gene promoter. Mol Cell Biol 1990; 10(7):3834-7.
[113]Kim YK, Lee AS. Identification of a 70-base-pair cell cycle regulatory unit within the promoter of the human thymidine kinase gene and its interaction with cellular factors. Mol Cell Biol 1991;11(4):2296-302.
[114]Kaelin WG, Jr., Krek W, Sellers WR, DeCaprio JA, Ajchenbaum F, Fuchs CS, et al. Expression cloning of a cDNA encoding a retinoblastoma-binding protein with E2F-like properties. Cell 1992;70(2):351-64.
[115]Farnham PJ, Slansky JE, Kollmar R. The role of E2F in the mammalian cell cycle. Biochim BiophysActa 1993; 1155(2):125-31.
[116]Slansky JE, Farnham PJ. Introduction to the E2F family:protein structure and gene regulation. CurrTop Microbiol Immunol 1996;208:1-30.
1 Ikeda M, Kawarabayashi T, Harigaya Y, et al. Motor impairment ameliorated by L-DOPA administration in human alpha-synuclein A30P+A53T transgenic mice with alpha-synuclein pathology. Brain Res,2008.
2 Ikeda M, Kawarabayashi T, Harigaya Y, et al.Motor impairment and aberrant production of neurochemicals in human alpha-synuclein A30P+A53T transgenic mice with alpha-synuclein pathology. Brain Res,2009; 1250:232-41.
3 Hirai Y, Fujita SC, Iwatsubo T, et al. Phosphorylated alpha-synuclein in normal mouse brain. FEBS Lett,2004; 572:227-32.
4 Takahashi M, Ko LW, Kulathingal J, et al. Oxidative stress-induced phosphorylation, degradation and aggregation of alpha-synuclein are linked to upregulated CK2 and cathepsin D. Eur J Neurosci,2007; 26:863-74.
5 Sakamoto M, Arawaka S, Hara S, et al.Contribution of endogenous G-protein-coupled receptor kinases to Ser129 phosphorylation of alpha-synuclein in HEK293 cells.Biochem Biophys Res Commun,2009;384(3):378-82.
6 Chau KY, Ching HL, Schapira AH, et al.Relationship between alpha synuclein phosphorylation, proteasomal inhibition and cell death:relevance to Parkinson's disease pathogenesis. J Neurochem,2009; 110(3):1005-13.
7 Qing H, Wong W, McGeer EG, et al.Lrrk2 phosphorylates alpha synuclein at serine 129:Parkinson disease implications. Biochem Biophys Res Commun,2009;387(1):149-52.
8 Kasai T, Tokuda T, Yamaguchi N, et al. Cleavage of normal and pathological forms of alpha-synuclein by neurosin in vitro. Neurosci Lett,2008; 436:52-6.
9 Smith WW, Margolis RL, Li X, et al. Alpha-synuclein phosphorylation enhances eosinophilic cytoplasmic inclusion formation in SH-SY5Y cells. J Neurosci,2005; 25:5544-52.
10 Liu C, Fei E, Jia N, et al. Assembly of lysine 63-linked ubiquitin conjugates by phosphorylated alpha-synuclein implies Lewy body biogenesis. J Biol Chem,2007; 282:14558-66.
11 Sugeno N, Takeda A, Hasegawa T, et al. Serine 129 phosphorylation of alpha-synuclein induces unfolded protein response-mediated cell death. J Biol Chem,2008; 283:23179-88.
12 Chen L, Feany MB. Alpha-synuclein phosphorylation controls neurotoxicity and inclusion formation in a Drosophila model of Parkinson disease. Nat Neurosci,2005; 8:657-63.
13 Kim EJ, Sung JY, Lee HJ, et al. DyrklA phosphorylates alpha-synuclein and enhances intracellular inclusion formation. J Biol Chem,2006; 281:33250-7.
14 Paleologou KE, Schmid AW, Rospigliosi CC, et al. Phosphorylation at Ser-129 but not the phosphomimics S129E/D inhibits the fibrillation of alpha-synuclein. J Biol Chem,2008; 283:16895-905.
15 Gorbatyuk OS, Li S, Sullivan LF, et al. The phosphorylation state of Ser-129 in human alpha-synuclein determines neurodegeneration in a rat model of Parkinson disease. Proc Natl Acad Sci U S A,2008; 105:763-8.
16 Wu B, Liu Q, Duan C, et al.Phosphorylation of alpha-synuclein upregulates tyrosine hydroxylase activity in MN9D cells. Acta Histochem,2009.
17 Assayag K, Yakunin E, Loeb V, et al. Polyunsaturated fatty acids induce alpha-synuclein-related pathogenic changes in neuronal cells. Am J Pathol,2007; 171:2000-11.
18 Gao HM, Kotzbauer PT, Uryu K, et al. Neuroinflammation and oxidation/nitration of alpha-synuclein linked to dopaminergic neurodegeneration. J Neurosci,2008; 28:7687-98.
19 Andrekopoulos C, Zhang H, Joseph J, et al. Bicarbonate enhances alpha-synuclein oligomerization and nitration:intermediacy of carbonate radical anion and nitrogen dioxide radical. Biochem J,2004; 378:435-47.
20 Shavali S, Combs CK, Ebadi M. Reactive macrophages increase oxidative stress and alpha-synuclein nitration during death of dopaminergic neuronal cells in co-culture:relevance to Parkinson's disease. Neurochem Res,2006; 31:85-94.
21 Hodara R, Norris EH, Giasson BI, et al. Functional consequences of alpha-synuclein tyrosine nitration:diminished binding to lipid vesicles and increased fibril formation. J Biol Chem,2004; 279:47746-53.
22 Reynolds AD, Glanzer JG, Kadiu I, et al. Nitrated alpha-synuclein-activated microglial profiling for Parkinson's disease. J Neurochem,2008; 104(6):1504-25.
23 Reynolds AD, Kadiu I, Garg SK, et al. Nitrated alpha-synuclein and microglial neuroregulatory activities J Neuroimmune Pharmacol,2008;3(2):59-74.
24 Yamin G, Uversky VN, Fink AL. Nitration inhibits fibrillation of human alpha-synuclein in vitro by formation of soluble oligomers. FEBS Lett,2003; 542: 147-52.
25 Xia Q, Liao L, Cheng D, et al. Proteomic identification of novel proteins associated with Lewy bodies. Front Biosci,2008; 13:3850-6.
26 Lee FK, Wong AK, Lee YW, et al.The role of ubiquitin linkages on alpha-synuclein induced-toxicity in a Drosophila model of Parkinson's disease.J Neurochem,2009;110(1):208-19.
27 Monti B, Polazzi E, Batti L, et al.Alpha-synuclein protects cerebellar granule neurons against 6-hydroxydopamine-induced death.,J Neurochem,2007; 103(2):518-30.
28 Rott R, Szargel R, Haskin J, et al. Monoubiquitylation of alpha-synuclein by seven in absentia homolog (SIAH) promotes its aggregation in dopaminergic cells. J Biol Chem,2008; 283:3316-28.
29 Lee JT, Wheeler TC, Li L, et al. Ubiquitination of alpha-synuclein by Siah-1 promotes alpha-synuclein aggregation and apoptotic cell death. Hum Mol Genet,2008; 17:906-17.
[2]Gasser T. Genetics of Parkinson's disease. J Neurol 2001;248(10):833-40.
[3]Maroteaux L, Campanelli JT, Scheller RH. Synuclein:a neuron-specific protein localized to the nucleus and presynaptic nerve terminal. J Neurosci 1988;8(8):2804-15.
[4]Conway KA, Harper JD, Lansbury PT, Jr. Fibrils formed in vitro from alpha-synuclein and two mutant forms linked to Parkinson's disease are typical amyloid. Biochemistry 2000;39(10):2552-63.
[5]El-Agnaf OM, Jakes R, Curran MD, Wallace A. Effects of the mutations Ala30 to Pro and Ala53 to Thr on the physical and morphological properties of alpha-synuclein protein implicated in Parkinson's disease. FEBS Lett 1998;440(1-2):67-70.
[6]Bisaglia M, Schievano E, Caporale A, Peggion E, Mammi S. The 11-mer repeats of human alpha-synuclein in vesicle interactions and lipid composition discrimination:a cooperative role. Biopolymers 2006;84(3):310-6.
[7]Davidson WS, Jonas A, Clayton DF, George JM. Stabilization of alpha-synuclein secondary structure upon binding to synthetic membranes. J Biol Chem 1998;273(16):9443-9.
[8]Jao CC, Der-Sarkissian A, Chen J, Langen R. Structure of membrane-bound alpha-synuclein studied by site-directed spin labeling. Proc Natl Acad Sci U S A 2004; 101(22):8331-6.
[9]Biere AL, Wood SJ, Wypych J, Steavenson S, Jiang Y, Anafi D, et al. Parkinson's disease-associated alpha-synuclein is more fibrillogenic than beta-and gamma-synuclein and cannot cross-seed its homologs. J Biol Chem 2000;275(44):34574-9.
[10]Perrin RJ, Woods WS, Clayton DF, George JM. Interaction of human alpha-Synuclein and Parkinson's disease variants with phospholipids. Structural analysis using site-directed mutagenesis. J Biol Chem 2000;275(44):34393-8.
[11]Weinreb PH, Zhen W, Poon AW, Conway KA, Lansbury PT, Jr. NACP, a protein implicated in Alzheimer's disease and learning, is natively unfolded. Biochemistry 1996;35(43):13709-15.
[12]Bussell R, Jr., Eliezer D. Effects of Parkinson's disease-linked mutations on the structure of lipid-associated alpha-synuclein. Biochemistry 2004;43(16):4810-8.
[13]Chandra S, Chen X, Rizo J, Jahn R, Sudhof TC. A broken alpha-helix in folded alpha-Synuclein. J Biol Chem 2003;278(17):15313-8.
[14]Trojanowski JQ, Lee VM. Parkinson's disease and related alpha-synucleinopathies are brain amyloidoses. Ann N Y Acad Sci 2003;991:107-10.
[15]Barghorn S, Davies P, Mandelkow E. Tau paired helical filaments from Alzheimer's disease brain and assembled in vitro are based on beta-structure in the core domain. Biochemistry 2004;43(6):1694-703.
[16]Giasson BI, Uryu K, Trojanowski JQ, Lee VM. Mutant and wild type human alpha-synucleins assemble into elongated filaments with distinct morphologies in vitro. J Biol Chem 1999;274(12):7619-22.
[17]Serpell LC, Berriman J, Jakes R, Goedert M, Crowther RA. Fiber diffraction of synthetic alpha-synuclein filaments shows amyloid-like cross-beta conformation. Proc Natl Acad Sci U S A 2000;97(9):4897-902.
[18]Kruger R, Kuhn W, Muller T, Woitalla D, Graeber M, Kosel S, et al. Ala30Pro mutation in the gene encoding alpha-synuclein in Parkinson's disease. Nat Genet 1998; 18(2):106-8.
[19]Polymeropoulos MH, Lavedan C, Leroy E, Ide SE, Dehejia A, Dutra A, et al. Mutation in the alpha-synuclein gene identified in families with Parkinson's disease. Science 1997;276(5321):2045-7.
[20]Zarranz JJ, Alegre J, Gomez-Esteban JC, Lezcano E, Ros R, Ampuero I, et al. The new mutation, E46K, of alpha-synuclein causes Parkinson and Lewy body dementia. Ann Neurol 2004;55(2):164-73.
[21]Greenbaum EA, Graves CL, Mishizen-Eberz AJ, Lupoli MA, Lynch DR, Englander SW, et al. The E46K mutation in alpha-synuclein increases amyloid fibril formation. J Biol Chem 2005;280(9):7800-7.
[22]Perrin RJ, Woods WS, Clayton DF, George JM. Exposure to long chain polyunsaturated fatty acids triggers rapid multimerization of synucleins. J Biol Chem 2001;276(45):41958-62.
[23]Li J, Uversky VN, Fink AL. Effect of familial Parkinson's disease point mutations A30P and A53T on the structural properties, aggregation, and fibrillation of human alpha-synuclein. Biochemistry 2001;40(38):11604-13.
[24]Smith WW, Margolis RL, Li X, Troncoso JC, Lee MK, Dawson VL, et al. Alpha-synuclein phosphorylation enhances eosinophilic cytoplasmic inclusion formation in SH-SY5Y cells. J Neurosci 2005;25(23):5544-52.
[25]Unger EL, Eve DJ, Perez XA, Reichenbach DK, Xu Y, Lee MK, et al. Locomotor hyperactivity and alterations in dopamine neurotransmission are associated with overexpression of A53T mutant human alpha-synuclein in mice. Neurobiol Dis 2006;21(2):431-43.
[26]Pandey N, Schmidt RE, Galvin JE. The alpha-synuclein mutation E46K promotes aggregation in cultured cells. Exp Neurol 2006; 197(2):515-20.
[27]Chen L, Feany MB. Alpha-synuclein phosphorylation controls neurotoxicity and inclusion formation in a Drosophila model of Parkinson disease. Nat Neurosci 2005;8(5):657-63.
[28]Choi W, Zibaee S, Jakes R, Serpell LC, Davletov B, Crowther RA, et al. Mutation E46K increases phospholipid binding and assembly into filaments of human alpha-synuclein. FEBS Lett 2004;576(3):363-8.
[29]Conway KA, Rochet JC, Bieganski RM, Lansbury PT, Jr. Kinetic stabilization of the alpha-synuclein protofibril by a dopamine-alpha-synuclein adduct. Science 2001;294(5545):1346-9.
[30]Lavedan C. The synuclein family. Genome Res 1998;8(9):871-80.
[31]Hsu LJ, Mallory M, Xia Y, Veinbergs I, Hashimoto M, Yoshimoto M, et al. Expression pattern of synucleins (non-Abeta component of Alzheimer's disease amyloid precursor protein/alpha-synuclein) during murine brain development. J Neurochem 1998;71(1):338-44.
[32]Bayer TA, Jakala P, Hartmann T, Egensperger R, Buslei R, Falkai P, et al. Neural expression profile of alpha-synuclein in developing human cortex. Neuroreport 1999;10(13):2799-803.
[33]Galvin JE, Schuck TM, Lee VM, Trojanowski JQ. Differential expression and distribution of alpha-, beta-, and gamma-synuclein in the developing human substantia nigra. Exp Neurol 2001;168(2):347-55.
[34]Petersen K, Olesen OF, Mikkelsen JD. Developmental expression of alpha-synuclein in rat hippocampus and cerebral cortex. Neuroscience 1999;91(2):651-9.
[35]Stefanis L, Kholodilov N, Rideout HJ, Burke RE, Greene LA. Synuclein-1 is selectively up-regulated in response to nerve growth factor treatment in PC 12 cells. J Neurochem 2001;76(4):1165-76.
[36]Vila M, Vukosavic S, Jackson-Lewis V, Neystat M, Jakowec M, Przedborski S. Alpha-synuclein up-regulation in substantia nigra dopaminergic neurons following administration of the parkinsonian toxin MPTP. J Neurochem 2000;74(2):721-9.
[37]Hashimoto M, Yoshimoto M, Sisk A, Hsu LJ, Sundsmo M, Kittel A, et al. NACP, a synaptic protein involved in Alzheimer's disease, is differentially regulated during megakaryocyte differentiation. Biochem Biophys Res Commun 1997;237(3):611-6.
[38]Cole NB, Murphy DD. The cell biology of alpha-synuclein:a sticky problem? Neuromolecular Med 2002; 1(2):95-109.
[39]Zhang L, Zhang C, Zhu Y, Cai Q, Chan P, Ueda K, et al. Semi-quantitative analysis of alpha-synuclein in subcellular pools of rat brain neurons:an immunogold electron microscopic study using a C-terminal specific monoclonal antibody. Brain Res 2008; 1244:40-52.
[40]Schneider BL, Seehus CR, Capowski EE, Aebischer P, Zhang SC, Svendsen CN. Over-expression of alpha-synuclein in human neural progenitors leads to specific changes in fate and differentiation. Hum Mol Genet 2007; 16(6):651-66.
[41]Andringa G, Du F, Chase TN, Bennett MC. Mapping of rat brain using the Synuclein-1 monoclonal antibody reveals somatodendritic expression of alpha-synuclein in populations of neurons homologous to those vulnerable to Lewy body formation in human synucleopathies. J Neuropathol Exp Neurol 2003;62(10):1060-75.
[42]Mori F, Tanji K, Yoshimoto M, Takahashi H, Wakabayashi K. Immunohistochemical comparison of alpha- and beta-synuclein in adult rat central nervous system. Brain Res 2002;941 (1-2):118-26.
[43]Specht CG, Tigaret CM, Rast GF, Thalhammer A, Rudhard Y, Schoepfer R. Subcellular localisation of recombinant alpha- and gamma-synuclein. Mol Cell Neurosci 2005;28(2):326-34.
[44]Takahashi M, Kanuka H, Fujiwara H, Koyama A, Hasegawa M, Miura M, et al. Phosphorylation of alpha-synuclein characteristic of synucleinopathy lesions is recapitulated in alpha-synuclein transgenic Drosophila. Neurosci Lett 2003;336(3):155-8.
[45]Masliah E, Rockenstein E, Veinbergs I, Mallory M, Hashimoto M, Takeda A, et al. Dopaminergic loss and inclusion body formation in alpha-synuclein mice: implications for neurodegenerative disorders. Science 2000;287(5456):1265-9.
[46]Goers J, Manning-Bog AB, McCormack AL, Millett IS, Doniach S, Di Monte DA, et al. Nuclear localization of alpha-synuclein and its interaction with histones. Biochemistry 2003;42(28):8465-71.
[47]McLean PJ, Ribich S, Hyman BT. Subcellular localization of alpha-synuclein in primary neuronal cultures:effect of missense mutations. J Neural Transm Suppl 2000(58):53-63.
[48]Yu S, Zuo X, Li Y, Zhang C, Zhou M, Zhang YA, et al. Inhibition of tyrosine hydroxylase expression in alpha-synuclein-transfected dopaminergic neuronal cells. Neurosci Lett 2004;367(1):34-9.
[49]Yu S, Li X, Liu G, Han J, Zhang C, Li Y, et al. Extensive nuclear localization of alpha-synuclein in normal rat brain neurons revealed by a novel monoclonal antibody. Neuroscience 2007;145(2):539-55.
[50]Leng Y, Chase TN, Bennett MC. Muscarinic receptor stimulation induces translocation of an alpha-synuclein oligomer from plasma membrane to a light vesicle fraction in cytoplasm. J Biol Chem 2001;276(30):28212-8.
[51]Hegde ML, Jagannatha Rao KS. Challenges and complexities of alpha-synuclein toxicity:new postulates in unfolding the mystery associated with Parkinson's disease. Arch Biochem Biophys 2003;418(2):169-78.
[52]Hegde ML, Rao KS. DNA induces folding in alpha-synuclein:understanding the mechanism using chaperone property of osmolytes. Arch Biochem Biophys 2007;464(1):57-69.
[53]Dev KK, Hofele K, Barbieri S, Buchman VL, van der Putten H. Part II: alpha-synuclein and its molecular pathophysiological role in neurodegenerative disease. Neuropharmacology 2003;45(1):14-44.
[54]Iwai A, Masliah E, Yoshimoto M, Ge N, Flanagan L, de Silva HA, et al. The precursor protein of non-A beta component of Alzheimer's disease amyloid is a presynaptic protein of the central nervous system. Neuron 1995;14(2):467-75.
[55]Spillantini MG, Schmidt ML, Lee VM, Trojanowski JQ, Jakes R, Goedert M. Alpha-synuclein in Lewy bodies. Nature 1997;388(6645):839-40.
[56]Wakabayashi K, Matsumoto K, Takayama K, Yoshimoto M, Takahashi H. NACP, a presynaptic protein, immunoreactivity in Lewy bodies in Parkinson's disease. Neurosci Lett 1997;239(1):45-8.
[57]Dingwall C, Sharnick SV, Laskey RA. A polypeptide domain that specifies migration of nucleoplasmin into the nucleus. Cell 1982;30(2):449-58.
[58]Kalderon D, Roberts BL, Richardson WD, Smith AE. A short amino acid sequence able to specify nuclear location. Cell 1984;39(3 Pt 2):499-509.
[59]Dingwall C, Laskey RA. Nuclear targeting sequences--a consensus? Trends Biochem Sci 1991;16(12):478-81.
[60]Dang CV, Lee WM. Identification of the human c-myc protein nuclear translocation signal. Mol Cell Biol 1988;8(10):4048-54.
[61]Zhou M, Liu H, Xu X, Zhou H, Li X, Peng C, et al. Identification of nuclear localization signal that governs nuclear import of BRD7 and its essential roles in inhibiting cell cycle progression. J Cell Biochem 2006;98(4):920-30.
[62]Hanaka H, Shimizu T, Izumi T. Nuclear-localization-signal-dependent and nuclear-export-signal-dependent mechanisms determine the localization of 5-lipoxygenase. Biochem J 2002;361(Pt 3):505-14.
[63]Antoine M, Reimers K, Wirz W, Gressner AM, Muller R, Kiefer P. Identification of an unconventional nuclear localization signal in human ribosomal protein S2. Biochem Biophys Res Commun 2005;335(l):146-53.
[64]Chan CK, Jans DA. Using nuclear targeting signals to enhance non-viral gene ransfer. Immunol Cell Biol 2002;80(2):119-30.
[65]Robbins J, Dilworth SM, Laskey RA, Dingwall C. Two interdependent basic domains in nucleoplasmin nuclear targeting sequence:identification of a class of bipartite nuclear targeting sequence. Cell 1991;64(3):615-23.
[66]Gasiorowski JZ, Dean DA. Mechanisms of nuclear transport and interventions. Adv Drug Deliv Rev 2003;55(6):703-16.
[67]Kotera I, Sekimoto T, Miyamoto Y, Saiwaki T, Nagoshi E, Sakagami H, et al. Importin alpha transports CaMKIV to the nucleus without utilizing importin beta. EmboJ2005;24(5):942-51.
[68]Jensen KF, Ohmstede CA, Fisher RS, Sahyoun N. Nuclear and axonal localization of Ca2+/calmodulin-dependent protein kinase type Gr in rat cerebellar cortex. Proc Natl Acad Sci U S A 1991;88(7):2850-3.
[69]Gorlich D, Kutay U. Transport between the cell nucleus and the cytoplasm. Annu Rev Cell Dev Biol 1999; 15:607-60.
[70]Komeili A, O'Shea EK. New perspectives on nuclear transport. Annu Rev Genet 2001;35:341-64.
[71]Nakielny S, Dreyfuss G. Import and export of the nuclear protein import receptor transportin by a mechanism independent of GTP hydrolysis. Curr Biol 1998;8(2):89-95.
[72]Ben-Ze'ev A, Shtutman M, Zhurinsky J. The integration of cell adhesion with gene expression:the role of beta-catenin. Exp Cell Res 2000;261(1):75-82.
[73]Fagotto F, Gluck U, Gumbiner BM. Nuclear localization signal-independent and importin/karyopherin-independent nuclear import of beta-catenin. Curr Biol 1998;8(4):181-90.
[74]Sachdev S, Bagchi S, Zhang DD, Mings AC, Hannink M. Nuclear import of IkappaBalpha is accomplished by a ran-independent transport pathway. Mol Cell Biol2000;20(5):1571-82.
[75]Sachdev S, Hoffmann A, Hannink M. Nuclear localization of IkappaB alpha is mediated by the second ankyrin repeat:the IkappaB alpha ankyrin repeats define a novel class of cis-acting nuclear import sequences. Mol Cell Biol 1998;18(5):2524-34.
[76]Hicks GR, Raikhel NV. Protein import into the nucleus:an integrated view. Annu Rev Cell Dev Biol 1995; 11:155-88.
[77]Kosugi S, Hasebe M, Entani T, Takayama S, Tomita M, Yanagawa H. Design of peptide inhibitors for the importin alpha/beta nuclear import pathway by activity-based profiling. Chem Biol 2008;15(9):940-9.
[78]Kontopoulos E, Parvin JD, Feany MB. Alpha-synuclein acts in the nucleus to inhibit histone acetylation and promote neurotoxicity. Hum Mol Genet 2006;15(20):3012-23.
[79]Baptista MJ, O'Farrell C, Daya S, Ahmad R, Miller DW, Hardy J, et al. Co-ordinate transcriptional regulation of dopamine synthesis genes by alpha-synuclein in human neuroblastoma cell lines. J Neurochem 2003;85(4):957-68.
[80]Lee MK, Stirling W, Xu Y, Xu X, Qui D, Mandir AS, et al. Human alpha-synuclein-harboring familial Parkinson's disease-linked Ala-53--> Thr mutation causes neurodegenerative disease with alpha-synuclein aggregation in transgenic mice. Proc Natl Acad Sci U S A 2002;99(13):8968-73.
[81]Cherny D, Hoyer W, Subramaniam V, Jovin TM. Double-stranded DNA stimulates the fibrillation of alpha-synuclein in vitro and is associated with the mature fibrils:an electron microscopy study. J Mol Biol 2004;344(4):929-38.
[82]Yamada M, Iwatsubo T, Mizuno Y, Mochizuki H. Overexpression of alpha-synuclein in rat substantia nigra results in loss of dopaminergic neurons, phosphorylation of alpha-synuclein and activation of caspase-9:resemblance to pathogenetic changes in Parkinson's disease. J Neurochem 2004;91 (2):451-61.
[83]Chesselet MF In vivo alpha-synuclein overexpression in rodents:a useful model of Parkinson's disease? ExpNeurol 2008;209(1):22-7.
[84]Lucking CB, Brice A. Alpha-synuclein and Parkinson's disease. Cell Mol Life Sci 2000;57(13-14):1894-908.
[85]Ostrerova N, Petrucelli L, Farrer M, Mehta N, Choi P, Hardy J, et al. alpha-Synuclein shares physical and functional homology with 14-3-3 proteins. J Neurosci 1999; 19(14):5782-91.
[86]Jenco JM, Rawlingson A, Daniels B, Morris AJ. Regulation of phospholipase D2: selective inhibition of mammalian phospholipase D isoenzymes by alpha-and beta-synucleins. Biochemistry 1998;37(14):4901-9.
[87]Klein J, Lindmar R, Loffelholz K. Muscarinic activation of phosphatidylcholine hydrolysis. Prog Brain Res 1996;109:201-8.
[88]Collas P, Husebye H, Alestrom P. The nuclear localization sequence of the SV40 T antigen promotes transgene uptake and expression in zebrafish embryo nuclei. Transgenic Res 1996;5(6):451-8.
[89]Xiao CY, Jans DA. An engineered site for protein kinase C flanking the SV40 large T-antigen NLS confers phorbol ester-inducible nuclear import. FEBS Lett 1998;436(3):313-7.
[90]Fukuda M, Gotoh I, Gotoh Y, Nishida E. Cytoplasmic localization of mitogen-activated protein kinase kinase directed by its NH2-terminal, leucine-rich short amino acid sequence, which acts as a nuclear export signal. J BiolChem 1996;271(33):20024-8.
[91]Nakamura S, Kawamoto Y, Nakano S, Akiguchi I, Kimura J. p35nck5a and cyclin-dependent kinase 5 colocalize in Lewy bodies of brains with Parkinson's disease. Acta Neuropathol 1997;94(2):153-7.
[92]Neystat M, Rzhetskaya M, Oo TF, Kholodilov N, Yarygina O, Wilson A, et al. Expression of cyclin-dependent kinase 5 and its activator p35 in models of induced apoptotic death in neurons of the substantia nigra in vivo. J Neurochem 2001;77(6):1611-25.
[93]Staropoli JF, McDermott C, Martinat C, Schulman B, Demireva E, Abeliovich A. Parkin is a component of an SCF-like ubiquitin ligase complex and protects postmitotic neurons from kainate excitotoxicity. Neuron 2003;37(5):735-49.
[94]Dauer W, Przedborski S. Parkinson's disease:mechanisms and models. Neuron 2003;39(6):889-909.
[95]Jordan-Sciutto KL, Dorsey R, Chalovich EM, Hammond RR, Achim CL. Expression patterns of retinoblastoma protein in Parkinson disease. J Neuropathol Exp Neurol 2003;62(1):68-74.
[96]El-Khodor BF, Oo TF, Kholodilov N, Burke RE. Ectopic expression of cell cycle markers in models of induced programmed cell death in dopamine neurons of the rat substantia nigra pars compacta. Exp Neurol 2003; 179(1):17-27.
[97]Hoglinger GU, Breunig JJ, Depboylu C, Rouaux C, Michel PP, Alvarez-Fischer D, et al. The pRb/E2F cell-cycle pathway mediates cell death in Parkinson's disease. Proc Natl Acad Sci U S A 2007;104(9):3585-90.
[98]Alvira D, Tajes M, Verdaguer E, de Arriba SG, Allgaier C, Matute C, et al. Inhibition of cyclin-dependent kinases is neuroprotective in l-methyl-4-phenylpyridinium-induced apoptosis in neurons. Neuroscience 2007;146(1):350-65.
[99]Lotharius J, O'Malley KL. The parkinsonism-inducing drug l-methyl-4-phenylpyridinium triggers intracellular dopamine oxidation. A novel mechanism of toxicity. J Biol Chem 2000;275(49):38581-8.
[100]Lee M, Hyun D, Halliwell B, Jenner P. Effect of the overexpression of wild-type or mutant alpha-synuclein on cell susceptibility to insult. J Neurochem 2001;76(4):998-1009.
[101]Wu F, Poon WS, Lu G, Wang A, Meng H, Feng L, et al. Alpha-synuclein knockdown attenuates MPP+induced mitochondrial dysfunction of SH-SY5Y cells. Brain Res 2009; 1292:173-9.
[102]Fountaine TM, Venda LL, Warrick N, Christian HC, Brundin P, Channon KM, et al. The effect of alpha-synuclein knockdown on MPP+toxicity in models of human neurons. Eur J Neurosci 2008;28(12):2459-73.
[103]Fountaine TM, Wade-Martins R. RNA interference-mediated knockdown of alpha-synuclein protects human dopaminergic neuroblastoma cells from MPP(+) toxicity and reduces dopamine transport. J Neurosci Res 2007;85(2):351-63.
[104]Pizarro JG, Junyent F, Verdaguer E, Jordan J, Beas-Zarate C, Pallas M, et al. Effects of MPP+on the molecular pathways involved in cell cycle control in B65 neuroblastoma cells. Pharmacol Res 2010;61(5):391-9.
[105]Bai J, Nakamura H, Ueda S, Kwon YW, Tanaka T, Ban S, et al. Proteasome-dependent degradation of cyclin D1 in 1-methyl-4-phenylpyridinium ion (MPP+)-induced cell cycle arrest. J Biol Chem 2004;279(37):38710-4.
[106]Gomez-Tortosa E, Newell K, Irizarry MC, Sanders JL, Hyman BT. alpha-Synuclein immunoreactivity in dementia with Lewy bodies: morphological staging and comparison with ubiquitin immunostaining. Acta Neuropathol 2000;99(4):352-7.
[107]Tanji K, Mori F, Imaizumi T, Yoshida H, Matsumiya T, Tamo W, et al. Upregulation of alpha-synuclein by lipopolysaccharide and interleukin-1 in human macrophages. Pathol Int 2002;52(9):572-7.
[108]Sangchot P, Sharma S, Chetsawang B, Porter J, Govitrapong P, Ebadi M. Deferoxamine attenuates iron-induced oxidative stress and prevents mitochondrial aggregation and alpha-synuclein translocation in SK-N-SH cells in culture. Dev Neurosci 2002;24(2-3):143-53.
[109]Lin WL, DeLucia MW, Dickson DW. Alpha-synuclein immunoreactivity in neuronal nuclear inclusions and neurites in multiple system atrophy. Neurosci Lett 2004;354(2):99-102.
[110]Hegde ML, Anitha S, Latha KS, Mustak MS, Stein R, Ravid R, et al. First evidence for helical transitions in supercoiled DNA by amyloid Beta Peptide (1-42) and aluminum:a new insight in understanding Alzheimer's disease. J Mol Neurosci 2004;22(1-2):19-31.
[111]Suram A, Hegde ML, Rao KS. A new evidence for DNA nicking property of amyloid beta-peptide (1-42):relevance to Alzheimer's disease. Arch Biochem Biophys 2007;463(2):245-52.
[112]Roehl HH, Conrad SE. Identification of a G1-S-phase-regulated region in the human thymidine kinase gene promoter. Mol Cell Biol 1990; 10(7):3834-7.
[113]Kim YK, Lee AS. Identification of a 70-base-pair cell cycle regulatory unit within the promoter of the human thymidine kinase gene and its interaction with cellular factors. Mol Cell Biol 1991;11(4):2296-302.
[114]Kaelin WG, Jr., Krek W, Sellers WR, DeCaprio JA, Ajchenbaum F, Fuchs CS, et al. Expression cloning of a cDNA encoding a retinoblastoma-binding protein with E2F-like properties. Cell 1992;70(2):351-64.
[115]Farnham PJ, Slansky JE, Kollmar R. The role of E2F in the mammalian cell cycle. Biochim BiophysActa 1993; 1155(2):125-31.
[116]Slansky JE, Farnham PJ. Introduction to the E2F family:protein structure and gene regulation. CurrTop Microbiol Immunol 1996;208:1-30.
1 Ikeda M, Kawarabayashi T, Harigaya Y, et al. Motor impairment ameliorated by L-DOPA administration in human alpha-synuclein A30P+A53T transgenic mice with alpha-synuclein pathology. Brain Res,2008.
2 Ikeda M, Kawarabayashi T, Harigaya Y, et al.Motor impairment and aberrant production of neurochemicals in human alpha-synuclein A30P+A53T transgenic mice with alpha-synuclein pathology. Brain Res,2009; 1250:232-41.
3 Hirai Y, Fujita SC, Iwatsubo T, et al. Phosphorylated alpha-synuclein in normal mouse brain. FEBS Lett,2004; 572:227-32.
4 Takahashi M, Ko LW, Kulathingal J, et al. Oxidative stress-induced phosphorylation, degradation and aggregation of alpha-synuclein are linked to upregulated CK2 and cathepsin D. Eur J Neurosci,2007; 26:863-74.
5 Sakamoto M, Arawaka S, Hara S, et al.Contribution of endogenous G-protein-coupled receptor kinases to Ser129 phosphorylation of alpha-synuclein in HEK293 cells.Biochem Biophys Res Commun,2009;384(3):378-82.
6 Chau KY, Ching HL, Schapira AH, et al.Relationship between alpha synuclein phosphorylation, proteasomal inhibition and cell death:relevance to Parkinson's disease pathogenesis. J Neurochem,2009; 110(3):1005-13.
7 Qing H, Wong W, McGeer EG, et al.Lrrk2 phosphorylates alpha synuclein at serine 129:Parkinson disease implications. Biochem Biophys Res Commun,2009;387(1):149-52.
8 Kasai T, Tokuda T, Yamaguchi N, et al. Cleavage of normal and pathological forms of alpha-synuclein by neurosin in vitro. Neurosci Lett,2008; 436:52-6.
9 Smith WW, Margolis RL, Li X, et al. Alpha-synuclein phosphorylation enhances eosinophilic cytoplasmic inclusion formation in SH-SY5Y cells. J Neurosci,2005; 25:5544-52.
10 Liu C, Fei E, Jia N, et al. Assembly of lysine 63-linked ubiquitin conjugates by phosphorylated alpha-synuclein implies Lewy body biogenesis. J Biol Chem,2007; 282:14558-66.
11 Sugeno N, Takeda A, Hasegawa T, et al. Serine 129 phosphorylation of alpha-synuclein induces unfolded protein response-mediated cell death. J Biol Chem,2008; 283:23179-88.
12 Chen L, Feany MB. Alpha-synuclein phosphorylation controls neurotoxicity and inclusion formation in a Drosophila model of Parkinson disease. Nat Neurosci,2005; 8:657-63.
13 Kim EJ, Sung JY, Lee HJ, et al. DyrklA phosphorylates alpha-synuclein and enhances intracellular inclusion formation. J Biol Chem,2006; 281:33250-7.
14 Paleologou KE, Schmid AW, Rospigliosi CC, et al. Phosphorylation at Ser-129 but not the phosphomimics S129E/D inhibits the fibrillation of alpha-synuclein. J Biol Chem,2008; 283:16895-905.
15 Gorbatyuk OS, Li S, Sullivan LF, et al. The phosphorylation state of Ser-129 in human alpha-synuclein determines neurodegeneration in a rat model of Parkinson disease. Proc Natl Acad Sci U S A,2008; 105:763-8.
16 Wu B, Liu Q, Duan C, et al.Phosphorylation of alpha-synuclein upregulates tyrosine hydroxylase activity in MN9D cells. Acta Histochem,2009.
17 Assayag K, Yakunin E, Loeb V, et al. Polyunsaturated fatty acids induce alpha-synuclein-related pathogenic changes in neuronal cells. Am J Pathol,2007; 171:2000-11.
18 Gao HM, Kotzbauer PT, Uryu K, et al. Neuroinflammation and oxidation/nitration of alpha-synuclein linked to dopaminergic neurodegeneration. J Neurosci,2008; 28:7687-98.
19 Andrekopoulos C, Zhang H, Joseph J, et al. Bicarbonate enhances alpha-synuclein oligomerization and nitration:intermediacy of carbonate radical anion and nitrogen dioxide radical. Biochem J,2004; 378:435-47.
20 Shavali S, Combs CK, Ebadi M. Reactive macrophages increase oxidative stress and alpha-synuclein nitration during death of dopaminergic neuronal cells in co-culture:relevance to Parkinson's disease. Neurochem Res,2006; 31:85-94.
21 Hodara R, Norris EH, Giasson BI, et al. Functional consequences of alpha-synuclein tyrosine nitration:diminished binding to lipid vesicles and increased fibril formation. J Biol Chem,2004; 279:47746-53.
22 Reynolds AD, Glanzer JG, Kadiu I, et al. Nitrated alpha-synuclein-activated microglial profiling for Parkinson's disease. J Neurochem,2008; 104(6):1504-25.
23 Reynolds AD, Kadiu I, Garg SK, et al. Nitrated alpha-synuclein and microglial neuroregulatory activities J Neuroimmune Pharmacol,2008;3(2):59-74.
24 Yamin G, Uversky VN, Fink AL. Nitration inhibits fibrillation of human alpha-synuclein in vitro by formation of soluble oligomers. FEBS Lett,2003; 542: 147-52.
25 Xia Q, Liao L, Cheng D, et al. Proteomic identification of novel proteins associated with Lewy bodies. Front Biosci,2008; 13:3850-6.
26 Lee FK, Wong AK, Lee YW, et al.The role of ubiquitin linkages on alpha-synuclein induced-toxicity in a Drosophila model of Parkinson's disease.J Neurochem,2009;110(1):208-19.
27 Monti B, Polazzi E, Batti L, et al.Alpha-synuclein protects cerebellar granule neurons against 6-hydroxydopamine-induced death.,J Neurochem,2007; 103(2):518-30.
28 Rott R, Szargel R, Haskin J, et al. Monoubiquitylation of alpha-synuclein by seven in absentia homolog (SIAH) promotes its aggregation in dopaminergic cells. J Biol Chem,2008; 283:3316-28.
29 Lee JT, Wheeler TC, Li L, et al. Ubiquitination of alpha-synuclein by Siah-1 promotes alpha-synuclein aggregation and apoptotic cell death. Hum Mol Genet,2008; 17:906-17.