基于微孔板核酸杂交酶联免疫吸附法检测DNA结合蛋白
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摘要
转录因子,也被称为序列特异性DNA结合蛋白,含有一个或多个DNA结合结构域(DBDs)。它们定位于细胞质中,在正常细胞功能状态下或受到特定因子诱导后,从细胞质进入细胞核,与基因组中的顺式作用元件诸如启动子、增强子、衰减子等发生特异性识别结合,组成基因转录机器,进而实现其调控基因表达的功能。因此,检测分析转录因子的表达及活化程度对我们了解细胞功能具有至关重要的作用。目前常用的检测技术主要建立在DNA和蛋白质相互作用这一层次,即运用含有顺式作用元件的DNA为探针,来分析转录因子的DNA结合活性。本课题“基于微孔板核酸杂交酶联免疫吸附法检测DNA结合蛋白”技术巧妙的利用在高严谨度洗脱条件下转录因子蛋白对与之特异性结合的双链核酸序列的保护作用,将对转录因子的检测转换为对相应的特异核酸序列的检测。该方法不需要复杂的仪器、繁琐的操作步骤和特异性抗体就可以对转录因子的表达及活化程度进行定量研究,而且可在较短时间内完成对转录因子的检测。我们以检测转录因子c-Jun的活性为模型来对该检测方法进行评价,在较短的时间内该方法可以准确的检测到3.5ng的转录因子c-Jun纯蛋白和0.625ug的Hela细胞核粗提取物,具有较高的灵敏度。而且该检测方法可以用于任何一种转录因子的检测。鉴于“基于微孔板核酸杂交酶联免疫吸附法”的快速、简便、高灵敏度和选择性,该检测方法可以应用到需要对转录因子的DNA结合活性进行定量,有效,大规模检测的领域和科学研究中,诸如细胞分子生物学中对多种处理的刺激应答筛选方面,进化生物学研究方面,环境监测方面和药物筛选方面等等。
Transcription factors, also known as sequence-specific DNA binding proteins,contain one or more DNA binding domains (DBDs). They are located in cytoplasm. In physiological condition or induced by a specific stimulus, transcription factors would be transported into the nucleus from cytoplasm, and then bind to trans-acting factors such as promoters, enhancers or attenuators, thereby up-regulating or down-regulating transcription of adjacent gene. Therefore, the determination of DNA binding activities of transcription factors is pivotal to our understanding of cellular functions. Currently used methods are all based on the interactions between DNA binding proteins and their counterparts, that is to say, use the trans-acting factors as probe to detect the DNA-binding activities of transcription factors. Because transcription factors can prevent specific double-stranded nucleic acids from being eluted to single-stranded in high stringency condition, herein, we detect the protected double-stranded nucleic acids to determine the DNA-binding activities of transcription factors indirectly. Microplate-nucleotide hybridization based ELISA-like assay did not require complex instruments and complicated procedure, and could be applied to multiplex detection without the usage of specific antibody in short time. We have used the method to demonstrate the DNA-binding activity of c-Jun, a well-known model of inductive transcriptional regulatory responses. But the method is easily adaptable for the study of any transcription factor. This allows rapid and accurate detection of as low as 3.5ng of purified c-Jun protein or 0.625ug crude cell nucleus extract. Owing to the rapidness, ease, high sensitivity and selectivity of the method, the microplate-nucleotide hybridization based ELISA-like assay may be used in various applications and research fields where quantitative, cost-effective and large-scale measurements of the DNA-binding activities of transcription factors are required, including screening of responses in multiple treatments in cellular and molecular biology, evolutionary biology, environmental monitoring, and drug discovery.
Transcription factors, also known as sequence-specific DNA binding proteins,contain one or more DNA binding domains (DBDs). They are located in cytoplasm. In physiological condition or induced by a specific stimulus, transcription factors would be transported into the nucleus from cytoplasm, and then bind to trans-acting factors such as promoters, enhancers or attenuators, thereby up-regulating or down-regulating transcription of adjacent gene. Therefore, the determination of DNA binding activities of transcription factors is pivotal to our understanding of cellular functions. Currently used methods are all based on the interactions between DNA binding proteins and their counterparts, that is to say, use the trans-acting factors as probe to detect the DNA-binding activities of transcription factors. Because transcription factors can prevent specific double-stranded nucleic acids from being eluted to single-stranded in high stringency condition, herein, we detect the protected double-stranded nucleic acids to determine the DNA-binding activities of transcription factors indirectly. Microplate-nucleotide hybridization based ELISA-like assay did not require complex instruments and complicated procedure, and could be applied to multiplex detection without the usage of specific antibody in short time. We have used the method to demonstrate the DNA-binding activity of c-Jun, a well-known model of inductive transcriptional regulatory responses. But the method is easily adaptable for the study of any transcription factor. This allows rapid and accurate detection of as low as 3.5ng of purified c-Jun protein or 0.625ug crude cell nucleus extract. Owing to the rapidness, ease, high sensitivity and selectivity of the method, the microplate-nucleotide hybridization based ELISA-like assay may be used in various applications and research fields where quantitative, cost-effective and large-scale measurements of the DNA-binding activities of transcription factors are required, including screening of responses in multiple treatments in cellular and molecular biology, evolutionary biology, environmental monitoring, and drug discovery.
引文
Bhattacharya, N., Sarno, A., Idler, I.S., et al. 2010. High-throughput detection of nuclear factor-kappaB activity using a sensitive oligo-based chemiluminescent enzyme-linked immunosorbent assay. International journal of cancer Journal international du cancer 127, 404-411.
Boon, E.M., Salas, J.E., and Barton, J.K. 2002. An electrical probe of protein-DNA interactions on DNA-modified surfaces. Nature biotechnology 20, 282-286.
Brenowitz, M., Senear, D.F., and Kingston, R.E. 2001. DNase I footprint analysis of protein-DNA binding. Current protocols in molecular biology / edited by Frederick M Ausubel [et al] Chapter 12, Unit 12 14.
Bulyk, M.L. 2006. DNA microarray technologies for measuring protein-DNA interactions. Current opinion in biotechnology 17, 422-430.
Cao, Y., and Karin, M. 2003. NF-kappaB in mammary gland development and breast cancer. Journal of mammary gland biology and neoplasia 8, 215-223.
Crinelli, R., Antonelli, A., Bianchi, M., et al. (2000). Selective inhibition of NF-kB activation and TNF-alpha production in macrophages by red blood cell-mediated delivery of dexamethasone. Blood cells, molecules & diseases 26, 211-222.
Ferrer, I., and Marti, E. 1998) Distribution of fibroblast growth factor receptor-1 (FGFR-1) and FGFR-3 in the hippocampus of patients with Alzheimer's disease. Neuroscience letters 240, 139-142.
Ferrer, I., Marti, E., Lopez, E., et al. 1998. NF-kB immunoreactivity is observed in association with beta A4 diffuse plaques in patients with Alzheimer's disease. Neuropathology and applied neurobiology 24, 271-277.
Galas, D.J., and Schmitz, A. 1978. DNAse footprinting: a simple method for the detection of protein-DNA binding specificity. Nucleic acids research 5, 3157-3170.
Garner, M.M., and Revzin, A. 1981. A gel electrophoresis method for quantifying the binding of proteins to specific DNA regions: application to components of the Escherichia coli lactose operon regulatory system. Nucleic acids research 9, 3047-3060.
Gupta, S.V., McGowen, R.M., Callewaert, D.M., et al. 2005. Quantitative chemiluescent immunoassay for NF-kappaB-DNA binding activity. Journal of immunoassay & immunochemistry 26, 125-143.
Hayden, M.S., and Ghosh, S. 2011. NF-kappaB in immunobiology. Cell research 21, 223-244.
He, H.J., Pires, R., Zhu, T.N., et al. 2007. Fluorescence resonance energy transfer-based method for detection of DNA binding activities of nuclear factor kappaB. BioTechniques 43, 93-98.
Iftode, C., Daniely, Y., and Borowiec, J.A. 1999. Replication protein A (RPA): the eukaryotic SSB. Critical reviews in biochemistry and molecular biology 34, 141-180.
Irwin, M.S., and Kaelin, W.G., Jr. 2001. Role of the newer p53 family proteins in malignancy. Apoptosis : an international journal on programmed cell death 6, 17-29.
Jagelska, E., Brazda, V., Pospisilova, S., et al. 2002. New ELISA technique for analysis of p53 protein/DNA binding properties. Journal of immunological methods 267, 227-235.
Jia, C.P., Zhong, X.Q., Hua, B., et al. 2009. Nano-ELISA for highly sensitive protein detection. Biosensors & bioelectronics 24, 2836-2841.
Jiao, X., Katiyar, S., Willmarth, N.E.,et al. 2010. c-Jun induces mammary epithelial cellular invasion and breast cancer stem cell expansion. The Journal of biological chemistry 285, 8218-8226.
Jin, S., Lu, D., Ye, S., et al. 2005. A simplified probe preparation for ELISA-based NF-kappaB activity assay. Journal of biochemical and biophysical methods 65, 20-29.
Joshi, B., Chakrabarty, A., Bruot, C., et al. 2010. DNA-WT1 protein interaction studied by surface-enhanced Raman spectroscopy. Analytical and bioanalytical chemistry 396, 1415-1421.
Kako, K., Wakamatsu, H., Hamada, T., et al. 1998. Examination of DNA-binding activity of neuronal transcription factors by electrophoretical mobility shift assay. Brain research Brain research protocols 2, 243-249.
Kitajima, I. 2007. Development of molecular pathology analysis testing which can be practiced in the hospital laboratory. The clinical application of a transcription factor activation test]. Rinsho byori The Japanese journal of clinical pathology 55, 262-271.
Knoll, E., and Heyduk, T. 2004. Unimolecular beacons for the detection of DNA-binding proteins. Analytical chemistry 76, 1156-1164.
Lahiri, D.K., and Ge, Y. 2000. Electrophoretic mobility shift assay for the detection of specific DNA-protein complex in nuclear extracts from the cultured cells and frozen autopsy human brain tissue. Brain research Brain research protocols 5, 257-265.
Latchman, D.S. 1997. Transcription factors: an overview. The international journal of biochemistry & cell biology 29, 1305-1312.
Lee, K.M., Kang, B.S., Lee, H.L., et al. 2004. Spinal NF-kB activation induces COX-2 upregulation and contributes to inflammatory pain hypersensitivity. The European journal of neuroscience 19, 3375-3381.
Lee, T.I., and Young, R.A. 2000. Transcription of eukaryotic protein-coding genes. Annual review of genetics 34, 77-137.
Libermann, T.A., and Zerbini, L.F. 2006. Targeting transcription factors for cancer gene therapy. Current gene therapy 6, 17-33.
Lin, A., and Karin, M. 2003. NF-kappaB in cancer: a marked target. Seminars in cancer biology 13, 107-114.
Lin, Y.L., Lai, Y.J., Tsai, N.M.,et al. 2010. A new microsphere-based immunoassay for measuring the activity of transcription factors. Biological procedures online 12, 9030.
Liu, B., Qin, Y., Wang, J., et al. 2011. Detection and comparison of protein-DNA interactions using DNA-BIND plate and horseradish peroxidase-based colorimetric assay. Analytical biochemistry 412, 111-113.
Lobe, C.G. .1992. Transcription factors and mammalian development. Current topics in developmental biology 27, 351-383.
Maeda, S., and Karin, M. 2003. Oncogene at last--c-Jun promotes liver cancer in mice. Cancer cell 3,102-104.
Menotta, M., Crinelli, R., Carloni, E., et al. 2010. Label-free quantification of activated NF-kappaB in biological samples by atomic force microscopy. Biosensors & bioelectronics 25, 2490-2496.
Meyyappan, M., Atadja, P.W., and Riabowol, K.T. 1996. Regulation of gene expression and transcription factor binding activity during cellular aging. Biological signals 5, 130-138.
Mitchell, P.J., and Tjian, R. 1989. Transcriptional regulation in mammalian cells by sequence-specific DNA binding proteins. Science 245, 371-378.
Myers, L.C., and Kornberg, R.D. 2000. Mediator of transcriptional regulation. Annual review of biochemistry 69, 729-749.
Nikolov, D.B., and Burley, S.K. 1997. RNA polymerase II transcription initiation: a structural view. Proceedings of the National Academy of Sciences of the United States of America 94, 15-22.
Oikawa, T. 2004. ETS transcription factors: possible targets for cancer therapy. Cancer science 95, 626-633.
Ou, L.J., Jin, P.Y., Chu, X., et al. 2010. Sensitive and visual detection of sequence-specific DNA-binding protein via a gold nanoparticle-based colorimetric biosensor. Analytical chemistry 82, 6015-6024.
Pabo, C.O., and Sauer, R.T. 1984. Protein-DNA recognition. Annual review of biochemistry 53, 293-321.
Pawson, T. 1993). Signal transduction--a conserved pathway from the membrane to the nucleus. Developmental genetics 14, 333-338.
Ptashne, M., and Gann, A. 1997. Transcriptional activation by recruitment. Nature 386, 569-577.
Raybaud-Diogene, H., Tetu, B., Morency, R., et al. 1996. p53 overexpression in head and neck squamous cell carcinoma: review of the literature. European journal of cancer Part B, Oral oncology 32B, 143-149.
Reese, J.C. 2003. Basal transcription factors. Current opinion in genetics & development 13, 114-118. Rosenau, C., Emery, D., Kaboord, B., et al. 2004. Development of a high-throughput plate-based chemiluminescent transcription factor assay. Journal of biomolecular screening 9, 334-342.
Sandman, K., Pereira, S.L., and Reeve, J.N. 1998. Diversity of prokaryotic chromosomal proteins and the origin of the nucleosome. Cellular and molecular life sciences : CMLS 54, 1350-1364.
Schutte, J., minna, J.D., and Birrer, M.J. 1989. Deregulated expression of human c-jun transforms primary rat embryo cells in cooperation with an activated c-Ha-ras gene and transforms rat-1a cells as a single gene. Proceedings of the National Academy of Sciences of the United States of America 86, 2257-2261.
Shamovsky, I., and Nudler, E. 2008. New insights into the mechanism of heat shock response activation. Cellular and molecular life sciences : CMLS 65, 855-861.
Spiegelman, B.M., and Heinrich, R. 2004. Biological control through regulated transcriptional coactivators. Cell 119, 157-167.
Tortosa, A., Lopez, E., and Ferrer, I. 1998. Bcl-2 and Bax protein expression in Alzheimer's disease. Acta neuropathologica 95, 407-412.
Viggiani, C.J., Aparicio, J.G., and Aparicio, O.M. 2009. ChIP-chip to analyze the binding of replicationproteins to chromatin using oligonucleotide DNA microarrays. Methods Mol Biol 521, 255-278.
Wang, J., Li, M.L., Hua, D.,et al. 2006a. Exonuclease-mediated ELISA-like assay for detecting DNA-binding activity of transcription factors: measurement of activated NF-kappaB. BioTechniques 41, 79-88, 90.
Wang, J.K., Li, J.L., Li, M.L.,et al. 2006b. Assay of DNA-binding proteins with a dsDNA-coupled plate. Clinical biochemistry 39, 167-175.
Wheaton, K., Atadja, P., and Riabowol, K. 1996. Regulation of transcription factor activity during cellular aging. Biochemistry and cell biology = Biochimie et biologie cellulaire 74, 523-534.
Wullaert, A., Bonnet, M.C., and Pasparakis, M. 2011. NF-kappaB in the regulation of epithelial homeostasis and inflammation. Cell research 21, 146-158.
Xin, W., Yun, K.J., Ricci, F., et al. 2004. MAP2K4/MKK4 expression in pancreatic cancer: genetic validation of immunohistochemistry and relationship to disease course. Clinical cancer research : an official journal of the American Association for Cancer Research 10, 8516-8520.
Xu, X., Zhao, Z., Qin, L.,et al. 2008. Fluorescence recovery assay for the detection of protein-DNA binding. Analytical chemistry 80, 5616-5621.
Zhang, Y., Pu, X., Shi, M., et al. 2007. c-Jun, a crucial molecule in metastasis of breast cancer and potential target for biotherapy. Oncology reports 18, 1207-1212.
Zhu, C., Byers, K.J., McCord, R.P., et al. 2009. High-resolution DNA-binding specificity analysis of yeast transcription factors. Genome research 19, 556-566.
Boon, E.M., Salas, J.E., and Barton, J.K. 2002. An electrical probe of protein-DNA interactions on DNA-modified surfaces. Nature biotechnology 20, 282-286.
Brenowitz, M., Senear, D.F., and Kingston, R.E. 2001. DNase I footprint analysis of protein-DNA binding. Current protocols in molecular biology / edited by Frederick M Ausubel [et al] Chapter 12, Unit 12 14.
Bulyk, M.L. 2006. DNA microarray technologies for measuring protein-DNA interactions. Current opinion in biotechnology 17, 422-430.
Cao, Y., and Karin, M. 2003. NF-kappaB in mammary gland development and breast cancer. Journal of mammary gland biology and neoplasia 8, 215-223.
Crinelli, R., Antonelli, A., Bianchi, M., et al. (2000). Selective inhibition of NF-kB activation and TNF-alpha production in macrophages by red blood cell-mediated delivery of dexamethasone. Blood cells, molecules & diseases 26, 211-222.
Ferrer, I., and Marti, E. 1998) Distribution of fibroblast growth factor receptor-1 (FGFR-1) and FGFR-3 in the hippocampus of patients with Alzheimer's disease. Neuroscience letters 240, 139-142.
Ferrer, I., Marti, E., Lopez, E., et al. 1998. NF-kB immunoreactivity is observed in association with beta A4 diffuse plaques in patients with Alzheimer's disease. Neuropathology and applied neurobiology 24, 271-277.
Galas, D.J., and Schmitz, A. 1978. DNAse footprinting: a simple method for the detection of protein-DNA binding specificity. Nucleic acids research 5, 3157-3170.
Garner, M.M., and Revzin, A. 1981. A gel electrophoresis method for quantifying the binding of proteins to specific DNA regions: application to components of the Escherichia coli lactose operon regulatory system. Nucleic acids research 9, 3047-3060.
Gupta, S.V., McGowen, R.M., Callewaert, D.M., et al. 2005. Quantitative chemiluescent immunoassay for NF-kappaB-DNA binding activity. Journal of immunoassay & immunochemistry 26, 125-143.
Hayden, M.S., and Ghosh, S. 2011. NF-kappaB in immunobiology. Cell research 21, 223-244.
He, H.J., Pires, R., Zhu, T.N., et al. 2007. Fluorescence resonance energy transfer-based method for detection of DNA binding activities of nuclear factor kappaB. BioTechniques 43, 93-98.
Iftode, C., Daniely, Y., and Borowiec, J.A. 1999. Replication protein A (RPA): the eukaryotic SSB. Critical reviews in biochemistry and molecular biology 34, 141-180.
Irwin, M.S., and Kaelin, W.G., Jr. 2001. Role of the newer p53 family proteins in malignancy. Apoptosis : an international journal on programmed cell death 6, 17-29.
Jagelska, E., Brazda, V., Pospisilova, S., et al. 2002. New ELISA technique for analysis of p53 protein/DNA binding properties. Journal of immunological methods 267, 227-235.
Jia, C.P., Zhong, X.Q., Hua, B., et al. 2009. Nano-ELISA for highly sensitive protein detection. Biosensors & bioelectronics 24, 2836-2841.
Jiao, X., Katiyar, S., Willmarth, N.E.,et al. 2010. c-Jun induces mammary epithelial cellular invasion and breast cancer stem cell expansion. The Journal of biological chemistry 285, 8218-8226.
Jin, S., Lu, D., Ye, S., et al. 2005. A simplified probe preparation for ELISA-based NF-kappaB activity assay. Journal of biochemical and biophysical methods 65, 20-29.
Joshi, B., Chakrabarty, A., Bruot, C., et al. 2010. DNA-WT1 protein interaction studied by surface-enhanced Raman spectroscopy. Analytical and bioanalytical chemistry 396, 1415-1421.
Kako, K., Wakamatsu, H., Hamada, T., et al. 1998. Examination of DNA-binding activity of neuronal transcription factors by electrophoretical mobility shift assay. Brain research Brain research protocols 2, 243-249.
Kitajima, I. 2007. Development of molecular pathology analysis testing which can be practiced in the hospital laboratory. The clinical application of a transcription factor activation test]. Rinsho byori The Japanese journal of clinical pathology 55, 262-271.
Knoll, E., and Heyduk, T. 2004. Unimolecular beacons for the detection of DNA-binding proteins. Analytical chemistry 76, 1156-1164.
Lahiri, D.K., and Ge, Y. 2000. Electrophoretic mobility shift assay for the detection of specific DNA-protein complex in nuclear extracts from the cultured cells and frozen autopsy human brain tissue. Brain research Brain research protocols 5, 257-265.
Latchman, D.S. 1997. Transcription factors: an overview. The international journal of biochemistry & cell biology 29, 1305-1312.
Lee, K.M., Kang, B.S., Lee, H.L., et al. 2004. Spinal NF-kB activation induces COX-2 upregulation and contributes to inflammatory pain hypersensitivity. The European journal of neuroscience 19, 3375-3381.
Lee, T.I., and Young, R.A. 2000. Transcription of eukaryotic protein-coding genes. Annual review of genetics 34, 77-137.
Libermann, T.A., and Zerbini, L.F. 2006. Targeting transcription factors for cancer gene therapy. Current gene therapy 6, 17-33.
Lin, A., and Karin, M. 2003. NF-kappaB in cancer: a marked target. Seminars in cancer biology 13, 107-114.
Lin, Y.L., Lai, Y.J., Tsai, N.M.,et al. 2010. A new microsphere-based immunoassay for measuring the activity of transcription factors. Biological procedures online 12, 9030.
Liu, B., Qin, Y., Wang, J., et al. 2011. Detection and comparison of protein-DNA interactions using DNA-BIND plate and horseradish peroxidase-based colorimetric assay. Analytical biochemistry 412, 111-113.
Lobe, C.G. .1992. Transcription factors and mammalian development. Current topics in developmental biology 27, 351-383.
Maeda, S., and Karin, M. 2003. Oncogene at last--c-Jun promotes liver cancer in mice. Cancer cell 3,102-104.
Menotta, M., Crinelli, R., Carloni, E., et al. 2010. Label-free quantification of activated NF-kappaB in biological samples by atomic force microscopy. Biosensors & bioelectronics 25, 2490-2496.
Meyyappan, M., Atadja, P.W., and Riabowol, K.T. 1996. Regulation of gene expression and transcription factor binding activity during cellular aging. Biological signals 5, 130-138.
Mitchell, P.J., and Tjian, R. 1989. Transcriptional regulation in mammalian cells by sequence-specific DNA binding proteins. Science 245, 371-378.
Myers, L.C., and Kornberg, R.D. 2000. Mediator of transcriptional regulation. Annual review of biochemistry 69, 729-749.
Nikolov, D.B., and Burley, S.K. 1997. RNA polymerase II transcription initiation: a structural view. Proceedings of the National Academy of Sciences of the United States of America 94, 15-22.
Oikawa, T. 2004. ETS transcription factors: possible targets for cancer therapy. Cancer science 95, 626-633.
Ou, L.J., Jin, P.Y., Chu, X., et al. 2010. Sensitive and visual detection of sequence-specific DNA-binding protein via a gold nanoparticle-based colorimetric biosensor. Analytical chemistry 82, 6015-6024.
Pabo, C.O., and Sauer, R.T. 1984. Protein-DNA recognition. Annual review of biochemistry 53, 293-321.
Pawson, T. 1993). Signal transduction--a conserved pathway from the membrane to the nucleus. Developmental genetics 14, 333-338.
Ptashne, M., and Gann, A. 1997. Transcriptional activation by recruitment. Nature 386, 569-577.
Raybaud-Diogene, H., Tetu, B., Morency, R., et al. 1996. p53 overexpression in head and neck squamous cell carcinoma: review of the literature. European journal of cancer Part B, Oral oncology 32B, 143-149.
Reese, J.C. 2003. Basal transcription factors. Current opinion in genetics & development 13, 114-118. Rosenau, C., Emery, D., Kaboord, B., et al. 2004. Development of a high-throughput plate-based chemiluminescent transcription factor assay. Journal of biomolecular screening 9, 334-342.
Sandman, K., Pereira, S.L., and Reeve, J.N. 1998. Diversity of prokaryotic chromosomal proteins and the origin of the nucleosome. Cellular and molecular life sciences : CMLS 54, 1350-1364.
Schutte, J., minna, J.D., and Birrer, M.J. 1989. Deregulated expression of human c-jun transforms primary rat embryo cells in cooperation with an activated c-Ha-ras gene and transforms rat-1a cells as a single gene. Proceedings of the National Academy of Sciences of the United States of America 86, 2257-2261.
Shamovsky, I., and Nudler, E. 2008. New insights into the mechanism of heat shock response activation. Cellular and molecular life sciences : CMLS 65, 855-861.
Spiegelman, B.M., and Heinrich, R. 2004. Biological control through regulated transcriptional coactivators. Cell 119, 157-167.
Tortosa, A., Lopez, E., and Ferrer, I. 1998. Bcl-2 and Bax protein expression in Alzheimer's disease. Acta neuropathologica 95, 407-412.
Viggiani, C.J., Aparicio, J.G., and Aparicio, O.M. 2009. ChIP-chip to analyze the binding of replicationproteins to chromatin using oligonucleotide DNA microarrays. Methods Mol Biol 521, 255-278.
Wang, J., Li, M.L., Hua, D.,et al. 2006a. Exonuclease-mediated ELISA-like assay for detecting DNA-binding activity of transcription factors: measurement of activated NF-kappaB. BioTechniques 41, 79-88, 90.
Wang, J.K., Li, J.L., Li, M.L.,et al. 2006b. Assay of DNA-binding proteins with a dsDNA-coupled plate. Clinical biochemistry 39, 167-175.
Wheaton, K., Atadja, P., and Riabowol, K. 1996. Regulation of transcription factor activity during cellular aging. Biochemistry and cell biology = Biochimie et biologie cellulaire 74, 523-534.
Wullaert, A., Bonnet, M.C., and Pasparakis, M. 2011. NF-kappaB in the regulation of epithelial homeostasis and inflammation. Cell research 21, 146-158.
Xin, W., Yun, K.J., Ricci, F., et al. 2004. MAP2K4/MKK4 expression in pancreatic cancer: genetic validation of immunohistochemistry and relationship to disease course. Clinical cancer research : an official journal of the American Association for Cancer Research 10, 8516-8520.
Xu, X., Zhao, Z., Qin, L.,et al. 2008. Fluorescence recovery assay for the detection of protein-DNA binding. Analytical chemistry 80, 5616-5621.
Zhang, Y., Pu, X., Shi, M., et al. 2007. c-Jun, a crucial molecule in metastasis of breast cancer and potential target for biotherapy. Oncology reports 18, 1207-1212.
Zhu, C., Byers, K.J., McCord, R.P., et al. 2009. High-resolution DNA-binding specificity analysis of yeast transcription factors. Genome research 19, 556-566.