p100蛋白与U5-116蛋白的相互结合作用
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摘要
目的:
真核基因的表达过程,包括DNA转录、pre-mRNA剪接、翻译等步骤。其中DNA转录和pre-mRNA剪接是两个重要环节,它们都要通过复杂的蛋白复合物来执行功能,涉及到蛋白-DNA、蛋白-RNA和蛋白-蛋白之间的相互作用。大量研究表明基因转录和pre-mRNA剪接并非两个独立的过程,多种蛋白具有参与转录和剪接调控的双重功能,从而将这两个过程紧密地连接起来,因此这些蛋白被称cross-talk蛋白。对于这些蛋白的了解有助于转录和剪接机制的深入探讨。人类p100蛋白是一个关键的转录调控子,能通过在启动子特异性激活因子和基本转录机器之间形成连接从而促进基因转录。HCA(hydrophobic cluster analysis,疏水簇分析)法发现p100蛋白是由4个重复的SN-lik(Staphylococcal nuclease-like,葡萄球菌核酸酶类似)功能片段和随后位于蛋白C末端Tudor-SN(TSN)功能片段组成。大量的研究发现p100蛋白作为一种共激活因子参与基因转录调控,这种作用主要借助其重复的SN样功能片段完成。前期我们课题组采用非变性胶电泳观察剪接体复合物体外组装的情况。观察发现p100蛋白TSN片段参与并促进剪接体的形成。但p100蛋白的TSN功能片段参与并促进pre-mRNA剪接加工过程的具体分子机制还不明确。
Pre-mRNA的剪接加工是在一个大的蛋白复合物—剪接体中进行的,而剪接体中重要的组分就是U5snRNP。U5-116蛋白因为能够直接与U5 snRNA结合,因而组成U5snRNP的核心,是U5的特异蛋白。本研究的主要目的在于研究p100蛋白,U5-116蛋白之间特异的结合方式和结合位点,通过蛋白—蛋白相互作用研究探讨p100蛋白的TSN功能片段参与并促进Pre-mRNA剪接加工过程的分子机制,为随后p100蛋白功能的研究奠定基础。
方法:
本课题分三部分进行研究,第一部分为确定能与p100蛋白,U5-116蛋白结合的hPrp8蛋白功能片段。首先根据hPrp8蛋白的结构特点构建hPrp8蛋白不同结构域的GST融合蛋白表达质粒。利用GST pull down方法用GST-hPrp8片段融合蛋白钓取体外翻译带有~(35)S标记的p100蛋白,放射自显影观察结果,寻找hPrp8的功能部位。同样方法研究U5-116蛋白与hPrp8蛋白结合的功能片段。
第二部分是p100蛋白与U5-116蛋白的结合。首先构建pEGFP-CI-U5-116重组质粒,利用CO-IP(Co-Immunoprecipitation,免疫共沉淀)技术研究p100蛋白与U5-116蛋白在细胞内的结合情况。再构建pERFP-CI-p100重组质粒,利用激光共聚焦显微镜研究p100蛋白与U5-116蛋白细胞内定位情况。
第三部分为了进一步确定p100蛋白与U5-116蛋白结合的功能片段。使用GST融合蛋白钓取法和Western Blot技术,用p100蛋白SN片段和TSN片段的GST融合蛋白(GST空载作为阴性对照)钓取细胞内过表达的U5-116蛋白,Western Blot方法检测。
结果:
第一部分hPrp8蛋白与p100蛋白,U5-116蛋白相互结合的功能片段研究结果显示:p100蛋白不能与hPrp8蛋白2.1片段结合,能与2.2片段在体外直接结合。U5-116蛋白能与hPrp8蛋白的2.1,2.2片段在体外结合。
第二部分成功构建了以下重组质粒:①pEGFP-CI-U5-116②pERFP-CI-p100。过表达的p100蛋白与U5-116蛋白之间可以发生共沉淀,表明两者细胞内可以稳定结合。利用激光共聚焦显微镜研究p100蛋白与U5-116蛋白定位情况。在转染后的细胞内观察到两者定位基本一致,主要分布于胞浆。
第三部分p100蛋白与U5-116蛋白在细胞外结合的研究结果显示p100蛋白的TSN片段能够与U5-116蛋白结合而SN片段不能与其结合。
结论:
本课题研究结果表明p100蛋白和U5-116蛋白都能与hPrp8蛋白体外直接结合,但结合部位不同。p100蛋白能够与U5-116蛋白在细胞内,外均可结合。二者之间的结合是由p100-TSN片段介导的。
Objective:
Eukaryotic gene expression process is consisting of DNA transcription, pre-mRNA splicing and translation.In this processes transcription and pre-mRNA splicing are the key processes and both of them require multi-component complex to function,concluding of protein-DNA,protein-RNA and protein-protein.Transcription and splicing are not independent events.They are functionally coordinated.Some proteins have transcription and splicing function,so they are called cross-talk protein. Studying these cross-talk proteins will help us known much about transcription and splicing.Human p100 protein is a novel transcriptional coactivator.HCA(hydrophobic cluster analysis)show that p100 consists of staphylococcal nuclease(SN)-like and Tudor(TSN)domains.The SN-like domains have been shown to function in transcription,but the function of TSN domain in the assembly of snRNP complexes and pre-mRNA splicing process.
Pre-mRNA splicing is happened in a big protein complex -splicosome,in which there is U5 snRNP specific proteins,such as U5-116.The purpose of this study was to validate the hypothesis that p100 protein functions in pre- mRNA splicing,and to reveal Splicing processes molecule mechanism.
Methods:
In this report,there are three parts.First,GST gene fusion proteins with various fragments of hPrp8 were used to pull down in vitro translated full length p100 or U5-116 labeled with~(35)S.Western Blot experiments were also used to confirm the specific interaction domains of hPrp8.
In the second part,interactions of p100 and U5-116 proteins in cells.We constructed the recombinant plasmids of pEGFP-CI-U5-116 and pERFP-CI-p100. The two recombinant plasmids are used to studies in cells.We coimmunoprecipitation of pl00 and U5-116 proteins to confirm the interaction between them.We also studied the colocalization of p100 and U5-116 proteins by immunofluorescence microscopy.
In the last part,identify the domain of p100 protein interacts with U5-116.GST gene fusions with various fragments of p100 were used to pull down U5-116 protein in cells to identify the interaction domains of p100.
Results:
In the first part,we found that p100 protein can bind to hPrp8 domain 2.2,not hPrp8 domain 2.1;U5-116 can bind to hPrp8 domain 2.1,2.2.
In the second part,we constructed the following recombinant plasmids successfully:①pEGFP-CI-U5-116②pERFP-CI-p100.Coimmunoprecipitation and Western Blot experiments also confirmed the interaction of over-express two proteins in cell.And we observed the colocalization of p100 protein and U5-116 protein by immunofluorescence microscopy.
In the third part,p100-TSN fragment stably interacts with U5-116 protein.
Conclusion:
Our present study provides following views:p100 protein,U5-116 protein can binds to hPrp8 protein,but function domains of hPrp8 are different.p100 protein can interacts U5-116 protein.p100-TSN stably binds to U5-116 protein.
真核基因的表达过程,包括DNA转录、pre-mRNA剪接、翻译等步骤。其中DNA转录和pre-mRNA剪接是两个重要环节,它们都要通过复杂的蛋白复合物来执行功能,涉及到蛋白-DNA、蛋白-RNA和蛋白-蛋白之间的相互作用。大量研究表明基因转录和pre-mRNA剪接并非两个独立的过程,多种蛋白具有参与转录和剪接调控的双重功能,从而将这两个过程紧密地连接起来,因此这些蛋白被称cross-talk蛋白。对于这些蛋白的了解有助于转录和剪接机制的深入探讨。人类p100蛋白是一个关键的转录调控子,能通过在启动子特异性激活因子和基本转录机器之间形成连接从而促进基因转录。HCA(hydrophobic cluster analysis,疏水簇分析)法发现p100蛋白是由4个重复的SN-lik(Staphylococcal nuclease-like,葡萄球菌核酸酶类似)功能片段和随后位于蛋白C末端Tudor-SN(TSN)功能片段组成。大量的研究发现p100蛋白作为一种共激活因子参与基因转录调控,这种作用主要借助其重复的SN样功能片段完成。前期我们课题组采用非变性胶电泳观察剪接体复合物体外组装的情况。观察发现p100蛋白TSN片段参与并促进剪接体的形成。但p100蛋白的TSN功能片段参与并促进pre-mRNA剪接加工过程的具体分子机制还不明确。
Pre-mRNA的剪接加工是在一个大的蛋白复合物—剪接体中进行的,而剪接体中重要的组分就是U5snRNP。U5-116蛋白因为能够直接与U5 snRNA结合,因而组成U5snRNP的核心,是U5的特异蛋白。本研究的主要目的在于研究p100蛋白,U5-116蛋白之间特异的结合方式和结合位点,通过蛋白—蛋白相互作用研究探讨p100蛋白的TSN功能片段参与并促进Pre-mRNA剪接加工过程的分子机制,为随后p100蛋白功能的研究奠定基础。
方法:
本课题分三部分进行研究,第一部分为确定能与p100蛋白,U5-116蛋白结合的hPrp8蛋白功能片段。首先根据hPrp8蛋白的结构特点构建hPrp8蛋白不同结构域的GST融合蛋白表达质粒。利用GST pull down方法用GST-hPrp8片段融合蛋白钓取体外翻译带有~(35)S标记的p100蛋白,放射自显影观察结果,寻找hPrp8的功能部位。同样方法研究U5-116蛋白与hPrp8蛋白结合的功能片段。
第二部分是p100蛋白与U5-116蛋白的结合。首先构建pEGFP-CI-U5-116重组质粒,利用CO-IP(Co-Immunoprecipitation,免疫共沉淀)技术研究p100蛋白与U5-116蛋白在细胞内的结合情况。再构建pERFP-CI-p100重组质粒,利用激光共聚焦显微镜研究p100蛋白与U5-116蛋白细胞内定位情况。
第三部分为了进一步确定p100蛋白与U5-116蛋白结合的功能片段。使用GST融合蛋白钓取法和Western Blot技术,用p100蛋白SN片段和TSN片段的GST融合蛋白(GST空载作为阴性对照)钓取细胞内过表达的U5-116蛋白,Western Blot方法检测。
结果:
第一部分hPrp8蛋白与p100蛋白,U5-116蛋白相互结合的功能片段研究结果显示:p100蛋白不能与hPrp8蛋白2.1片段结合,能与2.2片段在体外直接结合。U5-116蛋白能与hPrp8蛋白的2.1,2.2片段在体外结合。
第二部分成功构建了以下重组质粒:①pEGFP-CI-U5-116②pERFP-CI-p100。过表达的p100蛋白与U5-116蛋白之间可以发生共沉淀,表明两者细胞内可以稳定结合。利用激光共聚焦显微镜研究p100蛋白与U5-116蛋白定位情况。在转染后的细胞内观察到两者定位基本一致,主要分布于胞浆。
第三部分p100蛋白与U5-116蛋白在细胞外结合的研究结果显示p100蛋白的TSN片段能够与U5-116蛋白结合而SN片段不能与其结合。
结论:
本课题研究结果表明p100蛋白和U5-116蛋白都能与hPrp8蛋白体外直接结合,但结合部位不同。p100蛋白能够与U5-116蛋白在细胞内,外均可结合。二者之间的结合是由p100-TSN片段介导的。
Objective:
Eukaryotic gene expression process is consisting of DNA transcription, pre-mRNA splicing and translation.In this processes transcription and pre-mRNA splicing are the key processes and both of them require multi-component complex to function,concluding of protein-DNA,protein-RNA and protein-protein.Transcription and splicing are not independent events.They are functionally coordinated.Some proteins have transcription and splicing function,so they are called cross-talk protein. Studying these cross-talk proteins will help us known much about transcription and splicing.Human p100 protein is a novel transcriptional coactivator.HCA(hydrophobic cluster analysis)show that p100 consists of staphylococcal nuclease(SN)-like and Tudor(TSN)domains.The SN-like domains have been shown to function in transcription,but the function of TSN domain in the assembly of snRNP complexes and pre-mRNA splicing process.
Pre-mRNA splicing is happened in a big protein complex -splicosome,in which there is U5 snRNP specific proteins,such as U5-116.The purpose of this study was to validate the hypothesis that p100 protein functions in pre- mRNA splicing,and to reveal Splicing processes molecule mechanism.
Methods:
In this report,there are three parts.First,GST gene fusion proteins with various fragments of hPrp8 were used to pull down in vitro translated full length p100 or U5-116 labeled with~(35)S.Western Blot experiments were also used to confirm the specific interaction domains of hPrp8.
In the second part,interactions of p100 and U5-116 proteins in cells.We constructed the recombinant plasmids of pEGFP-CI-U5-116 and pERFP-CI-p100. The two recombinant plasmids are used to studies in cells.We coimmunoprecipitation of pl00 and U5-116 proteins to confirm the interaction between them.We also studied the colocalization of p100 and U5-116 proteins by immunofluorescence microscopy.
In the last part,identify the domain of p100 protein interacts with U5-116.GST gene fusions with various fragments of p100 were used to pull down U5-116 protein in cells to identify the interaction domains of p100.
Results:
In the first part,we found that p100 protein can bind to hPrp8 domain 2.2,not hPrp8 domain 2.1;U5-116 can bind to hPrp8 domain 2.1,2.2.
In the second part,we constructed the following recombinant plasmids successfully:①pEGFP-CI-U5-116②pERFP-CI-p100.Coimmunoprecipitation and Western Blot experiments also confirmed the interaction of over-express two proteins in cell.And we observed the colocalization of p100 protein and U5-116 protein by immunofluorescence microscopy.
In the third part,p100-TSN fragment stably interacts with U5-116 protein.
Conclusion:
Our present study provides following views:p100 protein,U5-116 protein can binds to hPrp8 protein,but function domains of hPrp8 are different.p100 protein can interacts U5-116 protein.p100-TSN stably binds to U5-116 protein.
引文
[1]Yang J,Tuuli Va" lineva,Jingxin Hong,et al.Transcriptional co-activator protein p100 interacts with snRNP proteins and facilitates the assembly of thespliceosome[J].Nucleic Acids Research,2007,35[13]:4485-4494
[2]Orphanides G,Reinberg D.A unified theory of gene expression[J].Cell,2002,108(4):439-451
[3]Will CL,Luhrmann R.Spliceosome structure and function in RNA world.Gestland RF,Cech TR,Atkins JF,eds.(Cold Spring Harbor,NY:Cold Spring Harbor Laboratory Press),369-400
[4]Karia MN.On the importance of being co-transcriptional[J].Cell Sci,2002,115(Pt20):3865-3871
[5]Wessagowit V,Nalla VK,Rogan PK,et al.Normal and abnormal mechanisms of gene splicing and relevance to inherited skin diseases[J].J Dermatol Sci,2005,42(2):73-84
[6]Mordes D,Luo X,Kar A,et al.Pre-mRNA splicing and retinitis pigmentosa[J].Mol Vis,2006,12:1259-1271
[7]Faustino NA,Cooper TA.Pre-mRNA splicing and human disease[J].Genes Dev,2003,17(4):419-437
[8]Bentley DL.Rules of engagement:co-transcriptional recruitment of pre-mRNA processing factors[J].Curr Opin Cell Biol,2005,17(3):251-256
[9]Hartmuth,K.,Vornlocher,H.P.and Luhrmann,R.Tobramycin affinity tag purification of spliceosomes[J].MethodsMol.Biol.,2004,257,47-64
[10]Rappsilber,J.,Ryder,U.,Lamond,A.I.and Mann,M.Large-scale proteomic analysis of the human spliceosome.Genome Res.,2002 12,1231-1245
[11]Zhou,Z.,Licklider,L.J.,Gygi,S.P.and Reed,R.Comprehensive proteomic analys-is of the human spliceosome.Nature,2002,419,182-185
[12]Bres V,Gomes N,Pickle L,et al.A human splicing factor,SKIP,associates with P-TEFb and enhances transcription elongation by HIV-1 Tat[J].Genes Dev,2005,19(10):1211-1226
[13]Nagai K,Yamaguchi T,Takami T,et al.SKIP modifies gene expression by affecting both transcription and splicing[J].Biochem Biophy Res Commu,2004,316(2):512-517
[14]Lin KT,Lu RM,Tarn WY.The WW domain-containing proteins interact with the early spliceosome and participate in pre-mRNA splicing in vivo[J].Mol Cell Biol,2004,24(20):9176-9185
[15]Jurica MS,Moore MJ.Pre-mRNA splicing:awash in a sea of proteins[J].Mol Cell,2003,12(1):5-14
[16]Staley JP,Guthrie C.Mechanical devices of the spliceosome:motors,clocks,springs,and things[J].Cell,1998,92(3):315-326
[17]Valadkhan S.snRNAs as the catalysts of pre-mRNA splicing[J].Curr Opin Chem,2005,9(6):603-608
[18]Will CL,Luhrmann R.Spliceosomal UsnRNP biogenesis,structure and function[J].Curr Opin Cell Biol,2001,13(3):290-301
[19]Graveley BR.Sorting out the complexity of SR protein functions[J].RNA,2000,6(9):1197-1211
[20]Tong,X.,Drapkin,R.,Yalamanchili,R.,Mosialos,G.and Kieff,E.The Epstein-Barr virus nuclear protein 2 acidic domainforms a complex with a novel cellular coactivator that can interactwith TFIIE[J].Mol.Cell.Biol.,1995,15,4735-4744.
[21]Qulle FW,Shimoda K,Thierfeider W,et al.Cloning of murine Stat6 and human Stat6:Stat proteins that are tyrosine phosphorylated in responses to IL-4 and IL-3 but are not required for mitogenesis[J].Mol Cell Biol,1995,15(6):3336-3343
[22]Wei D,Le x,Zheng L.et al.Stat3 activation regulate the expression of vascular endothelial growth factor and human pancreatic cancer angrogenesis and metst-asis[J].Ongene,2003,22(3):319-329
[23]Yang J,Alttomaki S,Pesu M,et al.Identification of p100 as a coactivator for STAT6 that bridges STAT6 with RNA polymerase Ⅱ[J].EMBO,2002,21(18):4950-4958
[24]Vlineva T,Yang J,Palovuori R.et al.The transcriptional co-activator protein p100 recruits histone acetyltransferase activity to STAT6 and mediates interaction[J].14989-14996
[25]Leverson JD,Koskinen PJ,et al.Pim-1 kinase and p100 cooperate to enhance c-Myb activity[J].Mol Cell,1998,2(4):417-425
[26]Marieke AT and Eric JS.Equine arteritis virus non-structural prtein 1,assential factor for viral subgenomic Mrna synthesis,interacts with the cellular transcri ption co-factor plOO[J].Journal of General Virology,2003,84,2317-2322
[27]Gregory RI,Chendrimada TP,Cooch N,et al.Human PISC couples micro RNA biogenesis and posttanscriptional gene silencing[J].Cell,2005,123:631-640
[28]Kim K,Lee YS,Carthew R.Conversion of pre-RISC to holo-RIAX by Ago2 during asembly of RNAi complexes[J].RNA,2007,13:22-29
[29]Connell MA,Keegan LP.Drosha versus ADAR:wrangling over pri-miRNA [J].Nat.Structu.Mol.Biol,2006,13:3-4
[30]Amy A,Rene FK,Scott M.et al.A micrococcal nuclease homologue in RNAi effector complexes [J].Nature,2003,(425):411-414
[31]Yang W,Chendrimada RP,Wang Q,et al.Modulation of microRNA processing and expression through RNA editing by ADARdeaminases[J].Nat.Struct.Mol.Biol,2006,13:13-21
[32]Callebaut I,Mornon JR The human EBNA-2 coactivator p100:multidomain organization and relationship to the staphylococcal nuclease fold and to the Tudor protein involved in Drosophila melanogaster development [J].Biochem J,1997,321(Pt1):125-132
[33]Strasswimmer J,Lorson CL,Breiding DE,et al.Identification of survival motor neuron as a transcriptional activator-binding protein [J].Hum Mol Genet,1999,8(7):1219-1226
[34]Girard C,Neel H,Bertrand E,et al.Depletion of SMN by RNA interference in HeLa cells induces defects in Caja1 body formation [J].Nucleic Acids Res,2006,34(10):2925-2932
[35]Golembe TJ,Yong J,DreyfussG.Specific sequence features,recognized by the SMN complex,identify snRNAs and determine their fate as snRNPs [J].Mol Cell Biol,2005,25 (24):10989-11004
[36]Tong X,Drapkin R,Yalamanchili R,Mosialos G and Kieff E.Epstein-Barr virus nuclear protein 2 acidic domain forms a complex with a novel cellular coactivator that can interact with TFFE [J].Mol Cell Biol.1995,15:4735-4744
[37]Ponting CP.Tudor domains in proteins that interact with RNA [J].Trends iochem Sci,1997,22(2):51-52
[38]Jie Yang,Saara AittomaEki,Marko Pesu,et al.Identification of p100 as acoactiv-ator for STAT6 that bridges STAT6 with RNA polymerase Ⅱ [J].EMBO,2002,21(18):4950-4958
[39]Callebaut I,Mornon JP.The human EBNA-2 coactivator p100:multidomain organization and relationship to the staphylococcal nuclease fold and to the tudor protein involved in Drosophila melanogaster development[J].Biochem,1997,321(Pt1):125-132
[40]Tuuli Valineva,Jie Yang,Riitta Palovuori,and Olli Silvennoinen.The Transcriptional Co-activator Protein p100 Recruits Histone Acetyltransferase Activity to STAT6 and Mediates Interaction between the CREB-binding Protein and STAT6[J].Biological chemistry,2005,280(15):14989-14996
[41]Pesu,M.,Aittomaki,S.,Valineva,T.,and Silvennoinen,O.PU.1 is required for transcriptional activation of the Stat6 response element in the lgepsilon promoter [J]Eur.J.Immunol,2003,33 (6):1727-1735
[42]Shankaranarayanan,P.,Chaitidis,P.,Kuhn,H.,and Nigam,S.Acetylation by histone acetyltransterase CREB-binding protein/p300 of STAT6 is required for transcriptional activation of the 15-lipoxygenase-1 gene[J].Biol.Chem,2001,276(46):42753-42760
[43]Tuuli Valineva,Jie Yang and Olli Silvennoinen.Characterization of RNA helicase A as component of STAT6-dependent enhanceosome [J].Nucleic Acids Research,2006,34 (14):3938-3946
[44]Grosschedl,R.Higher-order nucleoprotein complexes in transcription:Analogies with site-specific recombination [J].Curr.Opin.Cell Biol.,1995,7(3):362-370
[45]Thanos,D.and Maniatis,T.Virus induction of human IFN beta gene expression requires the assembly of an enhanceosome [J].Cell,1995,83(7):1091-1100
[46]Carey,M.The enhanceosome and transcriptional synergy [J].Cell,1998,92(1):5-8
[47]Ptashne,M.and Gann,A.Transcriptional activation by recruitment [J].Nature,1997,386(6625):569-577
[48]Naar,A.M.,Lemon,B.D.and Tjian,R.Transcriptional coactivator complexes[J].Annu.Rev.Biochem.,2001,70:475-501
[49]Zhang,S.and Grosse,F.Multiple functions of nuclear DNA helicase Ⅱ(RNA helicaseA)in nucleic acid metabolism [J].Acta Biochim.Biophys.Sin.(Shanghai),2004,36(3)177-183
[50]Lee,C.G.and Hurwitz,J.A new RNA helicase isolated from HeLa cells that cata-lytically translocates in the 30 to 50 direction [J].Biol.Chem.,1992,267(7):4398-4407
[51]Zhang,S.and Grosse,F.Nuclear DNA helicase Ⅱ unwinds both DNA and RNA [J].Biochemistry,1994,33(13):3906-3912
[52]Nakajima,T.,Uchida,C,Anderson,S.F.,et al.RNA helicase A mediates associate-on of CBP with RNA polymerase Ⅱ [J].Cell,1997,90(6):1107-1112
[53]Zhou,K.,Choe,K.T.,Zaidi,Z.,et al.RNA helicase A interacts with dsDNA and topoisomerase Ⅱ alpha[J].Nucleic Acids Res.,2003,31(9):2253-2260
[54]Anderson,S.F.,Schlegel,B.P.,Nakajima,T.,et al.BRCAl protein is linked to the RNA polymerase Ⅱ holoenzyme complex via RNA helicase A [J].Nat.Genet,1998,19(3):254-256
[55]Pellizzoni,L.,Charroux,B.,Rappsilber,J.,et al.A functional interaction between the survivalmotor neuron complex and RNA polymerase Ⅱ[J].Cell Biol.,2001,152(1):75-85
[56]Zhang,S.,Herrmann,C.and Grosse,F.Pre-mRNA and mRNA binding of humannuclear DNA helicase Ⅱ (RNA helicase A)[J],Cell.Sci.,1999,112(Pt7):1055-1064
[57]Lin KT,Lu RM,Tarn WY.The WW domain-containing proteins with the early spliceosome and participate in pre-mRNA splicing in vivo [J].Mol Cell Biol,2004,24(20):9176-9185
[58]Boon K L,Norman C M,Grainger J,et al.Prp8p dissection reveals domain structure and protein interaction sites [J].RNA,2006,12(2):198-205
[59]Turner IA,Norman CM,Churcher MJ,et al.Roles of the U5 snRNP in pliceosome dynamics and catalysis [J].Biochem Soc Trans,2004,32(Pt 6):928-931
[60]Bellare P,Kutach AK,Rines AK,et al.Ubiquitin binding by a variant Jabl/MPN domain in the essential pre-mRNA splicing factor Prp8p[J].RNA.2006,12(2):292-302
[61]Battels C,Urlaub H,Luhrmann R,et al.Mutagenesis Suggests Several Roles of Snu114p in Pre-mRNA Splicing[J].Biological chemistry,2003,278(30):2832.4-28334
[62]Dix I,Russell CS,O'Keefe RT,et al.Protein-RNA interactions in the U5 snRNP of Saccharomyces cerevisiae [J].RNA,1998,4(12):1675-1686
[63]Vidal VP,Verdone L,Mayes AE,et al.Characterization of U6 snRNA-protein interactions[J].RNA,1999,5(11):1470-1481
[64]Collins CA,Guthrie C.The question remains:Is the splicosome a ribozyme?[J].Nat Struct Biol,2000,7(10):850-854
[65]Grainger RJ,Beggs JD.Prp8 protein:At the heart of the spliceosome [J].RNA,2005,11(5):533-557
[66]Teigelkamp S,Whittaker E,Beggs JD.Interaction of the yeast splicing factor PRP8 with substrate RNA during both steps of splicing [J].Nucleic.Acids Res,1995,23 (3):320-326
[67]Pena V,Liu S,Bujnicki JM,et al.Structure of a multipartite protein-protein I nteraction domain in splicing factor prp8 and its link to retinitis pigmentosa[J].Mol Cell,2007,25(4):615-24
[68]Bartels C,Urlaub H,Luhrmann R,et al.Mutagenesis Suggests Several Roles of Snu114p in Pre-mRNA Splicing[J].Biological chemistry,2003,278(30):28324-2 8334
[69]Fabrizio P,Laggerbauer B,Lauber J,et al.An evolutionary conserved U5 snRNP-specific protein is a GTP-binding factor closely related to the ribosomal translocase EF-2[J].EMBO,1997,16 (13):4092-4106
[70]Patrizia Fabrizio,Bernhard Laggerbauer,Jiirgen Lauber,et al.An evolutionarily conserved U5 snRNP-specific protein is a GTP-binding factor closely related to theribosomal translocase EF-2 [J].EMBO,1997,16 (13):4092-4106
[71]Tamara J.Brenner and Christine Guthrie 1 Genetic Analysis Reveals a Role for the C Terminus of the Saccharomyces cerevisiae GTPase Snu114 during Spliceosome Activation [J].Genetics,2005,170:1063-1080
[72]Brenner TJ,Guthrie C.Assembly of Snu114 into U5 snRNP requires Prp8 and a functional GTPase domain [J].RNA,2006,12(5):862-71
[73]Bartels,C,Klatt,Luhrmann R,et al.The ribosomal translocase homologue Snu114p is involved in unwinding U4/U6 RNA during axtivation of the spliceosome [J].EMBO,2002,2(9):875-880
[74]Small E C,Leggett S R,Winans A A,et al.The EF-G-like GTPase Snu114p regulates spliceosome dynamics mediated by Brr2p,a DExD/H box ATPase[J].Mol Cel.2006,23(3):389-99
[75]Makarova OV,Makarov EM,Luhrmann R.The 65 and 110 kDa SR-related proteins of the U4/U6.U5 tri-snRNP are essential for the assembly of mature spliceosomes [J].EMBO,2001,20(10):2553-2563
[76]Raghunathan P,Guthrie C.RNA unwinding in U4/U6 snRNPs requires ATP hydrolysis and the DEIH-box splicing factor Brr2 [J].Current Biology 1998,8:847-855
[77]Bottnen C A,Schmidt H,Vogel S,et al.Norbert F.Kauter.Multiple genetic and biochemical interactions of Brr2,Prp8,Prp31,Prpl and Prp4 kinase suggest a function in the control of the activation of spliceosomes in Schizasacchanomyces pombe.[J].Curr Cenet,2005,48:151-161
[1]Gonsalvez G B,Tian L P,Ospina J K.Two distinct arginine methyltransferases are requiredfor biogenesis of Sm-class ribonucleoproteins [J].Cell Biology,2007,178 (5):733-740
[2]Newman,A.J.The role of U5 snRNP in pre-mRNA splicing[J].EMBO,1997,16(19):5797-5800
[3]Staley J P,Guthrie C.Mechanical devicesof the spliceosome:motors,cloks,springs,andthings[J].Cell,1998,92(3):315-326
[4]Grainger R J,Beggs J D.Prp8 protein:At the heart of the spliceosome [J].RNA,2005,11:533-557
[5]Turner I A,Norman C M,Churcher M J et al.Roles of the U5 snRNP in spliceo some dynamicsand catalysis[J].Biochemical Society,2004,32(6):928-931
[6]Pena V,Liu S,Bujnicki JM,et al.Structure of a multipartite protein-protein interaction domain in splicing factor prp8 and its link to retinitis pigmentosa[J].Mol Cell,2007,25(4):615-24
[7]Boon K L,Grainger RJ,Ehsani P,et al.Prp8 mutations that cause human retinitis pigmentosa lead to a U5 snRNP maturation defect in yeast[J].Nat Struct Mol Biol.,2007,14(11):1077-83
[8]Boon K L,Norman C M,Grainger J,et al.Prp8p dissection reveals domain structure and protein interaction sites [J].RNA,2006,12(2):198-205
[9]Bellare P,Kutach AK,Rines AK,et al.Ubiquitin binding by a variant Jab1/MPNdomain in the essential pre-mRNA splicing factor Prp8p[J].RNA.2006,12(2):292-302
[10]Bartels C,Urlaub H,Luhrmann R,et al.Mutagenesis Suggests Several Roles of Snu114p in Pre-mRNA Splicing[J].Biologicalchemistry,2003,278(30):28324-28334
[11]Fabrizio P,Laggerbauer B,Lauber J,et al.An evolutionarily conserved U5 snRNP-specific protein is a GTP-binding factor closely related to the ribosomal translocase EF-2[J].EMBO,1997,16(13):4092-4106
[12]Gottschalk,A,Neubauer,G.,Banroques,J.,et al.Identification by massspectro metry and functional analyses of novel proteins of the yeast tri-snRNP[J].E MBO,1999,18(16):4535-4548
[13]Tamara J.Brenner and Christine Guthrie 1.Genetic Analysis Reveals a Role for the C Terminus of the Saccharomyces cerevisiae GTPase Snu114 During Spliceosome Activation[J].Genetics,2005,170:1063-1080
[14]Brenner TJ,Guthrie C.Assembly of Snu114 into U5 snRNP requires Prp8 and a functional GTPase domain [J].RNA,2006,12(5):862-71
[15]Bartels,C,Klatt,Luhrmann R,et al.The ribosomal translocase homologue Snu114p is involved in unwinding U4/U6 RNA during axtivation of the spliceosome [J].EMBO,2002,2(9):875-880
[16]Small E C,Leggett S R,Winans A A,et al.The EF-G-like GTPase Snu114p regulates spliceosome dynamics mediated by Brr2p,a DExD/H box ATPase[J].Mol Cell.2006,23(3):389-99
[17]Makarova OV,Makarov EM,Luhrmann R.The 65 and 110 kDa SR-related proteins of the U4/U6.U5 tri-snRNP are essential for the assembly of mature spliceosomes[J].EMBO,2001,20(10):2553-2563
[18]Raghunathan P,Guthrie C.RNA unwinding in U4/U6 snRNPs requires ATP hydrolysis and the DEIH-box splicing factor Brr2[J].Current Biology 1998,8:847-855
[19]Bottnen C A,Schmidt H,Vogel S,et al.Norbert F.Kauter.Multiple genetic and biochemical interactions of Brr2,Prp8,Prp31,Prpl and Prp4 kinase suggest a function in the control of the activation of spliceosomes in Schizasacchanomyces pombe[J].Curr Cenet,2005,48:151-161
[20]Tsai R T,Tseng C K,Lee P J.Dynamic Interactions of Ntr1-Ntr2 with Prp43 and with U5 Governthe Recruitment of Prp43 To Mediate Spliceosome Disassembly [J].Molecular and cellular biology,2007,p.8027-8037
[21]Shen H,Green M R.A pathway of sequential arginine-serine-rich domain-splicing signal interactions during mammalian spliceosome assembly[J].Mol Cell,2004,16(3):363-373
[22]Ismaili N,Sha M,Gustafson E H,et al.The 100-kda U5 snRNP protein (hPrp28p)contacts the 5' splice site through its ATPase site[XJ.RNA,2001,7(2):182-193
[23]Mathew R,Hartmuth K,Mohlmann S,et al.Phosphorylation of human PRP28 by SRPK2 is required for integration of the U4/U6-U5 tri-snRNP into the spliceosome [J].Nat Struct Mol Biol,2008,15(5):435-43
[2]Orphanides G,Reinberg D.A unified theory of gene expression[J].Cell,2002,108(4):439-451
[3]Will CL,Luhrmann R.Spliceosome structure and function in RNA world.Gestland RF,Cech TR,Atkins JF,eds.(Cold Spring Harbor,NY:Cold Spring Harbor Laboratory Press),369-400
[4]Karia MN.On the importance of being co-transcriptional[J].Cell Sci,2002,115(Pt20):3865-3871
[5]Wessagowit V,Nalla VK,Rogan PK,et al.Normal and abnormal mechanisms of gene splicing and relevance to inherited skin diseases[J].J Dermatol Sci,2005,42(2):73-84
[6]Mordes D,Luo X,Kar A,et al.Pre-mRNA splicing and retinitis pigmentosa[J].Mol Vis,2006,12:1259-1271
[7]Faustino NA,Cooper TA.Pre-mRNA splicing and human disease[J].Genes Dev,2003,17(4):419-437
[8]Bentley DL.Rules of engagement:co-transcriptional recruitment of pre-mRNA processing factors[J].Curr Opin Cell Biol,2005,17(3):251-256
[9]Hartmuth,K.,Vornlocher,H.P.and Luhrmann,R.Tobramycin affinity tag purification of spliceosomes[J].MethodsMol.Biol.,2004,257,47-64
[10]Rappsilber,J.,Ryder,U.,Lamond,A.I.and Mann,M.Large-scale proteomic analysis of the human spliceosome.Genome Res.,2002 12,1231-1245
[11]Zhou,Z.,Licklider,L.J.,Gygi,S.P.and Reed,R.Comprehensive proteomic analys-is of the human spliceosome.Nature,2002,419,182-185
[12]Bres V,Gomes N,Pickle L,et al.A human splicing factor,SKIP,associates with P-TEFb and enhances transcription elongation by HIV-1 Tat[J].Genes Dev,2005,19(10):1211-1226
[13]Nagai K,Yamaguchi T,Takami T,et al.SKIP modifies gene expression by affecting both transcription and splicing[J].Biochem Biophy Res Commu,2004,316(2):512-517
[14]Lin KT,Lu RM,Tarn WY.The WW domain-containing proteins interact with the early spliceosome and participate in pre-mRNA splicing in vivo[J].Mol Cell Biol,2004,24(20):9176-9185
[15]Jurica MS,Moore MJ.Pre-mRNA splicing:awash in a sea of proteins[J].Mol Cell,2003,12(1):5-14
[16]Staley JP,Guthrie C.Mechanical devices of the spliceosome:motors,clocks,springs,and things[J].Cell,1998,92(3):315-326
[17]Valadkhan S.snRNAs as the catalysts of pre-mRNA splicing[J].Curr Opin Chem,2005,9(6):603-608
[18]Will CL,Luhrmann R.Spliceosomal UsnRNP biogenesis,structure and function[J].Curr Opin Cell Biol,2001,13(3):290-301
[19]Graveley BR.Sorting out the complexity of SR protein functions[J].RNA,2000,6(9):1197-1211
[20]Tong,X.,Drapkin,R.,Yalamanchili,R.,Mosialos,G.and Kieff,E.The Epstein-Barr virus nuclear protein 2 acidic domainforms a complex with a novel cellular coactivator that can interactwith TFIIE[J].Mol.Cell.Biol.,1995,15,4735-4744.
[21]Qulle FW,Shimoda K,Thierfeider W,et al.Cloning of murine Stat6 and human Stat6:Stat proteins that are tyrosine phosphorylated in responses to IL-4 and IL-3 but are not required for mitogenesis[J].Mol Cell Biol,1995,15(6):3336-3343
[22]Wei D,Le x,Zheng L.et al.Stat3 activation regulate the expression of vascular endothelial growth factor and human pancreatic cancer angrogenesis and metst-asis[J].Ongene,2003,22(3):319-329
[23]Yang J,Alttomaki S,Pesu M,et al.Identification of p100 as a coactivator for STAT6 that bridges STAT6 with RNA polymerase Ⅱ[J].EMBO,2002,21(18):4950-4958
[24]Vlineva T,Yang J,Palovuori R.et al.The transcriptional co-activator protein p100 recruits histone acetyltransferase activity to STAT6 and mediates interaction[J].14989-14996
[25]Leverson JD,Koskinen PJ,et al.Pim-1 kinase and p100 cooperate to enhance c-Myb activity[J].Mol Cell,1998,2(4):417-425
[26]Marieke AT and Eric JS.Equine arteritis virus non-structural prtein 1,assential factor for viral subgenomic Mrna synthesis,interacts with the cellular transcri ption co-factor plOO[J].Journal of General Virology,2003,84,2317-2322
[27]Gregory RI,Chendrimada TP,Cooch N,et al.Human PISC couples micro RNA biogenesis and posttanscriptional gene silencing[J].Cell,2005,123:631-640
[28]Kim K,Lee YS,Carthew R.Conversion of pre-RISC to holo-RIAX by Ago2 during asembly of RNAi complexes[J].RNA,2007,13:22-29
[29]Connell MA,Keegan LP.Drosha versus ADAR:wrangling over pri-miRNA [J].Nat.Structu.Mol.Biol,2006,13:3-4
[30]Amy A,Rene FK,Scott M.et al.A micrococcal nuclease homologue in RNAi effector complexes [J].Nature,2003,(425):411-414
[31]Yang W,Chendrimada RP,Wang Q,et al.Modulation of microRNA processing and expression through RNA editing by ADARdeaminases[J].Nat.Struct.Mol.Biol,2006,13:13-21
[32]Callebaut I,Mornon JR The human EBNA-2 coactivator p100:multidomain organization and relationship to the staphylococcal nuclease fold and to the Tudor protein involved in Drosophila melanogaster development [J].Biochem J,1997,321(Pt1):125-132
[33]Strasswimmer J,Lorson CL,Breiding DE,et al.Identification of survival motor neuron as a transcriptional activator-binding protein [J].Hum Mol Genet,1999,8(7):1219-1226
[34]Girard C,Neel H,Bertrand E,et al.Depletion of SMN by RNA interference in HeLa cells induces defects in Caja1 body formation [J].Nucleic Acids Res,2006,34(10):2925-2932
[35]Golembe TJ,Yong J,DreyfussG.Specific sequence features,recognized by the SMN complex,identify snRNAs and determine their fate as snRNPs [J].Mol Cell Biol,2005,25 (24):10989-11004
[36]Tong X,Drapkin R,Yalamanchili R,Mosialos G and Kieff E.Epstein-Barr virus nuclear protein 2 acidic domain forms a complex with a novel cellular coactivator that can interact with TFFE [J].Mol Cell Biol.1995,15:4735-4744
[37]Ponting CP.Tudor domains in proteins that interact with RNA [J].Trends iochem Sci,1997,22(2):51-52
[38]Jie Yang,Saara AittomaEki,Marko Pesu,et al.Identification of p100 as acoactiv-ator for STAT6 that bridges STAT6 with RNA polymerase Ⅱ [J].EMBO,2002,21(18):4950-4958
[39]Callebaut I,Mornon JP.The human EBNA-2 coactivator p100:multidomain organization and relationship to the staphylococcal nuclease fold and to the tudor protein involved in Drosophila melanogaster development[J].Biochem,1997,321(Pt1):125-132
[40]Tuuli Valineva,Jie Yang,Riitta Palovuori,and Olli Silvennoinen.The Transcriptional Co-activator Protein p100 Recruits Histone Acetyltransferase Activity to STAT6 and Mediates Interaction between the CREB-binding Protein and STAT6[J].Biological chemistry,2005,280(15):14989-14996
[41]Pesu,M.,Aittomaki,S.,Valineva,T.,and Silvennoinen,O.PU.1 is required for transcriptional activation of the Stat6 response element in the lgepsilon promoter [J]Eur.J.Immunol,2003,33 (6):1727-1735
[42]Shankaranarayanan,P.,Chaitidis,P.,Kuhn,H.,and Nigam,S.Acetylation by histone acetyltransterase CREB-binding protein/p300 of STAT6 is required for transcriptional activation of the 15-lipoxygenase-1 gene[J].Biol.Chem,2001,276(46):42753-42760
[43]Tuuli Valineva,Jie Yang and Olli Silvennoinen.Characterization of RNA helicase A as component of STAT6-dependent enhanceosome [J].Nucleic Acids Research,2006,34 (14):3938-3946
[44]Grosschedl,R.Higher-order nucleoprotein complexes in transcription:Analogies with site-specific recombination [J].Curr.Opin.Cell Biol.,1995,7(3):362-370
[45]Thanos,D.and Maniatis,T.Virus induction of human IFN beta gene expression requires the assembly of an enhanceosome [J].Cell,1995,83(7):1091-1100
[46]Carey,M.The enhanceosome and transcriptional synergy [J].Cell,1998,92(1):5-8
[47]Ptashne,M.and Gann,A.Transcriptional activation by recruitment [J].Nature,1997,386(6625):569-577
[48]Naar,A.M.,Lemon,B.D.and Tjian,R.Transcriptional coactivator complexes[J].Annu.Rev.Biochem.,2001,70:475-501
[49]Zhang,S.and Grosse,F.Multiple functions of nuclear DNA helicase Ⅱ(RNA helicaseA)in nucleic acid metabolism [J].Acta Biochim.Biophys.Sin.(Shanghai),2004,36(3)177-183
[50]Lee,C.G.and Hurwitz,J.A new RNA helicase isolated from HeLa cells that cata-lytically translocates in the 30 to 50 direction [J].Biol.Chem.,1992,267(7):4398-4407
[51]Zhang,S.and Grosse,F.Nuclear DNA helicase Ⅱ unwinds both DNA and RNA [J].Biochemistry,1994,33(13):3906-3912
[52]Nakajima,T.,Uchida,C,Anderson,S.F.,et al.RNA helicase A mediates associate-on of CBP with RNA polymerase Ⅱ [J].Cell,1997,90(6):1107-1112
[53]Zhou,K.,Choe,K.T.,Zaidi,Z.,et al.RNA helicase A interacts with dsDNA and topoisomerase Ⅱ alpha[J].Nucleic Acids Res.,2003,31(9):2253-2260
[54]Anderson,S.F.,Schlegel,B.P.,Nakajima,T.,et al.BRCAl protein is linked to the RNA polymerase Ⅱ holoenzyme complex via RNA helicase A [J].Nat.Genet,1998,19(3):254-256
[55]Pellizzoni,L.,Charroux,B.,Rappsilber,J.,et al.A functional interaction between the survivalmotor neuron complex and RNA polymerase Ⅱ[J].Cell Biol.,2001,152(1):75-85
[56]Zhang,S.,Herrmann,C.and Grosse,F.Pre-mRNA and mRNA binding of humannuclear DNA helicase Ⅱ (RNA helicase A)[J],Cell.Sci.,1999,112(Pt7):1055-1064
[57]Lin KT,Lu RM,Tarn WY.The WW domain-containing proteins with the early spliceosome and participate in pre-mRNA splicing in vivo [J].Mol Cell Biol,2004,24(20):9176-9185
[58]Boon K L,Norman C M,Grainger J,et al.Prp8p dissection reveals domain structure and protein interaction sites [J].RNA,2006,12(2):198-205
[59]Turner IA,Norman CM,Churcher MJ,et al.Roles of the U5 snRNP in pliceosome dynamics and catalysis [J].Biochem Soc Trans,2004,32(Pt 6):928-931
[60]Bellare P,Kutach AK,Rines AK,et al.Ubiquitin binding by a variant Jabl/MPN domain in the essential pre-mRNA splicing factor Prp8p[J].RNA.2006,12(2):292-302
[61]Battels C,Urlaub H,Luhrmann R,et al.Mutagenesis Suggests Several Roles of Snu114p in Pre-mRNA Splicing[J].Biological chemistry,2003,278(30):2832.4-28334
[62]Dix I,Russell CS,O'Keefe RT,et al.Protein-RNA interactions in the U5 snRNP of Saccharomyces cerevisiae [J].RNA,1998,4(12):1675-1686
[63]Vidal VP,Verdone L,Mayes AE,et al.Characterization of U6 snRNA-protein interactions[J].RNA,1999,5(11):1470-1481
[64]Collins CA,Guthrie C.The question remains:Is the splicosome a ribozyme?[J].Nat Struct Biol,2000,7(10):850-854
[65]Grainger RJ,Beggs JD.Prp8 protein:At the heart of the spliceosome [J].RNA,2005,11(5):533-557
[66]Teigelkamp S,Whittaker E,Beggs JD.Interaction of the yeast splicing factor PRP8 with substrate RNA during both steps of splicing [J].Nucleic.Acids Res,1995,23 (3):320-326
[67]Pena V,Liu S,Bujnicki JM,et al.Structure of a multipartite protein-protein I nteraction domain in splicing factor prp8 and its link to retinitis pigmentosa[J].Mol Cell,2007,25(4):615-24
[68]Bartels C,Urlaub H,Luhrmann R,et al.Mutagenesis Suggests Several Roles of Snu114p in Pre-mRNA Splicing[J].Biological chemistry,2003,278(30):28324-2 8334
[69]Fabrizio P,Laggerbauer B,Lauber J,et al.An evolutionary conserved U5 snRNP-specific protein is a GTP-binding factor closely related to the ribosomal translocase EF-2[J].EMBO,1997,16 (13):4092-4106
[70]Patrizia Fabrizio,Bernhard Laggerbauer,Jiirgen Lauber,et al.An evolutionarily conserved U5 snRNP-specific protein is a GTP-binding factor closely related to theribosomal translocase EF-2 [J].EMBO,1997,16 (13):4092-4106
[71]Tamara J.Brenner and Christine Guthrie 1 Genetic Analysis Reveals a Role for the C Terminus of the Saccharomyces cerevisiae GTPase Snu114 during Spliceosome Activation [J].Genetics,2005,170:1063-1080
[72]Brenner TJ,Guthrie C.Assembly of Snu114 into U5 snRNP requires Prp8 and a functional GTPase domain [J].RNA,2006,12(5):862-71
[73]Bartels,C,Klatt,Luhrmann R,et al.The ribosomal translocase homologue Snu114p is involved in unwinding U4/U6 RNA during axtivation of the spliceosome [J].EMBO,2002,2(9):875-880
[74]Small E C,Leggett S R,Winans A A,et al.The EF-G-like GTPase Snu114p regulates spliceosome dynamics mediated by Brr2p,a DExD/H box ATPase[J].Mol Cel.2006,23(3):389-99
[75]Makarova OV,Makarov EM,Luhrmann R.The 65 and 110 kDa SR-related proteins of the U4/U6.U5 tri-snRNP are essential for the assembly of mature spliceosomes [J].EMBO,2001,20(10):2553-2563
[76]Raghunathan P,Guthrie C.RNA unwinding in U4/U6 snRNPs requires ATP hydrolysis and the DEIH-box splicing factor Brr2 [J].Current Biology 1998,8:847-855
[77]Bottnen C A,Schmidt H,Vogel S,et al.Norbert F.Kauter.Multiple genetic and biochemical interactions of Brr2,Prp8,Prp31,Prpl and Prp4 kinase suggest a function in the control of the activation of spliceosomes in Schizasacchanomyces pombe.[J].Curr Cenet,2005,48:151-161
[1]Gonsalvez G B,Tian L P,Ospina J K.Two distinct arginine methyltransferases are requiredfor biogenesis of Sm-class ribonucleoproteins [J].Cell Biology,2007,178 (5):733-740
[2]Newman,A.J.The role of U5 snRNP in pre-mRNA splicing[J].EMBO,1997,16(19):5797-5800
[3]Staley J P,Guthrie C.Mechanical devicesof the spliceosome:motors,cloks,springs,andthings[J].Cell,1998,92(3):315-326
[4]Grainger R J,Beggs J D.Prp8 protein:At the heart of the spliceosome [J].RNA,2005,11:533-557
[5]Turner I A,Norman C M,Churcher M J et al.Roles of the U5 snRNP in spliceo some dynamicsand catalysis[J].Biochemical Society,2004,32(6):928-931
[6]Pena V,Liu S,Bujnicki JM,et al.Structure of a multipartite protein-protein interaction domain in splicing factor prp8 and its link to retinitis pigmentosa[J].Mol Cell,2007,25(4):615-24
[7]Boon K L,Grainger RJ,Ehsani P,et al.Prp8 mutations that cause human retinitis pigmentosa lead to a U5 snRNP maturation defect in yeast[J].Nat Struct Mol Biol.,2007,14(11):1077-83
[8]Boon K L,Norman C M,Grainger J,et al.Prp8p dissection reveals domain structure and protein interaction sites [J].RNA,2006,12(2):198-205
[9]Bellare P,Kutach AK,Rines AK,et al.Ubiquitin binding by a variant Jab1/MPNdomain in the essential pre-mRNA splicing factor Prp8p[J].RNA.2006,12(2):292-302
[10]Bartels C,Urlaub H,Luhrmann R,et al.Mutagenesis Suggests Several Roles of Snu114p in Pre-mRNA Splicing[J].Biologicalchemistry,2003,278(30):28324-28334
[11]Fabrizio P,Laggerbauer B,Lauber J,et al.An evolutionarily conserved U5 snRNP-specific protein is a GTP-binding factor closely related to the ribosomal translocase EF-2[J].EMBO,1997,16(13):4092-4106
[12]Gottschalk,A,Neubauer,G.,Banroques,J.,et al.Identification by massspectro metry and functional analyses of novel proteins of the yeast tri-snRNP[J].E MBO,1999,18(16):4535-4548
[13]Tamara J.Brenner and Christine Guthrie 1.Genetic Analysis Reveals a Role for the C Terminus of the Saccharomyces cerevisiae GTPase Snu114 During Spliceosome Activation[J].Genetics,2005,170:1063-1080
[14]Brenner TJ,Guthrie C.Assembly of Snu114 into U5 snRNP requires Prp8 and a functional GTPase domain [J].RNA,2006,12(5):862-71
[15]Bartels,C,Klatt,Luhrmann R,et al.The ribosomal translocase homologue Snu114p is involved in unwinding U4/U6 RNA during axtivation of the spliceosome [J].EMBO,2002,2(9):875-880
[16]Small E C,Leggett S R,Winans A A,et al.The EF-G-like GTPase Snu114p regulates spliceosome dynamics mediated by Brr2p,a DExD/H box ATPase[J].Mol Cell.2006,23(3):389-99
[17]Makarova OV,Makarov EM,Luhrmann R.The 65 and 110 kDa SR-related proteins of the U4/U6.U5 tri-snRNP are essential for the assembly of mature spliceosomes[J].EMBO,2001,20(10):2553-2563
[18]Raghunathan P,Guthrie C.RNA unwinding in U4/U6 snRNPs requires ATP hydrolysis and the DEIH-box splicing factor Brr2[J].Current Biology 1998,8:847-855
[19]Bottnen C A,Schmidt H,Vogel S,et al.Norbert F.Kauter.Multiple genetic and biochemical interactions of Brr2,Prp8,Prp31,Prpl and Prp4 kinase suggest a function in the control of the activation of spliceosomes in Schizasacchanomyces pombe[J].Curr Cenet,2005,48:151-161
[20]Tsai R T,Tseng C K,Lee P J.Dynamic Interactions of Ntr1-Ntr2 with Prp43 and with U5 Governthe Recruitment of Prp43 To Mediate Spliceosome Disassembly [J].Molecular and cellular biology,2007,p.8027-8037
[21]Shen H,Green M R.A pathway of sequential arginine-serine-rich domain-splicing signal interactions during mammalian spliceosome assembly[J].Mol Cell,2004,16(3):363-373
[22]Ismaili N,Sha M,Gustafson E H,et al.The 100-kda U5 snRNP protein (hPrp28p)contacts the 5' splice site through its ATPase site[XJ.RNA,2001,7(2):182-193
[23]Mathew R,Hartmuth K,Mohlmann S,et al.Phosphorylation of human PRP28 by SRPK2 is required for integration of the U4/U6-U5 tri-snRNP into the spliceosome [J].Nat Struct Mol Biol,2008,15(5):435-43