期刊文献+

与PIH1D1相互作用的蛋白分析

Analysis of the interaction proteins of PIH1D1
下载PDF
导出
摘要 目的分析细胞中与PIH1D1相互作用的蛋白。方法构建稳定表达FLAG-HA双标签标记的PIH1D1蛋白的HEK293T细胞株,利用FLAG-HA串联亲和纯化(TAP)双标签纯化实验,对目的条带进行质谱分析。结果成功构建稳定表达FLAG-HA双标签标记的PIH1D1细胞株。通过质谱分析得到了PIH1D1相互作用的蛋白数据,包括细胞质内RNA PolⅡ组装复合物成员RPAP3、UXT、PFD2和PFD6等,凋亡复合物成员MONAD/WDR92等,钙调蛋白信号通路中的PIP和CALM1以及代谢通路中的PKM和LCN1等。结论 PIH1D1与细胞中RPAP3、UXT、PFD2、PFD6、MONAD/WDR92、PIP、CALM1、PKM和LCN1等相互作用,提示PIH1D1可能参与细胞中RNApolⅡ组装、细胞凋亡、钙调蛋白通路等多种生理过程。 Objective To investigate HEK293T cell strain was constructed the binding proteins of PIH1 D1. using a FLAG-HA double-tagged Methods A PIHID1 stable-expressed PIHIDI expressing plasmid. After per- forming the FLAG-HA tandem affinity purification (TAP) , the mass spectrometry was carried out to analyze the interaction protein complex of PIH1D. Results The FLAG-HA double-tagged PIHID stable-expressed HEK293T cell strain was obtained. We found a number of potential binding proteins of PIHID1 by means of the mass spectrometry, including RPAP3, UXT, PFD2 and PFD6 which were involved in RNAP ]I assemble factory and MONAD/WDR92 which was able to promote apoptosis. Besides we found PIP and CALM1 belong- ing to calmodulin-Ca2+ complex, and PKM and LCN1 which were classic enzyme of metabolism as binding proteins of PIHID1 for the first time. Conclusions PIH1DI may participate in multiple biological processes including apoptosis, calmodulin-Ca(2 + ) signaling pathway, RNA polymerase II assembling, and metabo- lism regulation through interaction with a RPAP3, UXT, PFD2, PFD6, MONAD/WDR92, PIP, CALM1, PKM and LCN1.
作者 章元 张业
出处 《基础医学与临床》 CSCD 2016年第7期951-955,共5页 Basic and Clinical Medicine
基金 国家自然科学基金(31371305)
关键词 PIH1D1 串联亲和纯化 质谱 PIH1 D1 TAP mass spectrometric
  • 相关文献

参考文献8

  • 1Kakihara Y, Saeki M. The R2TP chaperone complex : its in-volvement in snoRNP assembly and tumorigenesis[ J]. Bio-mol Concepts, 2014, 5: 513-520.
  • 2Horejsi Z,Takai H, Adelman GA, et al. GK2 phospho-de-pendent binding of R2TP complex to TEL2 is essential formTOR and SMG1 stability [ J ]. Mol Cell, 2010,39:839-850.
  • 3Kakihara Y, Houry WA. The R2TP complex : discovery andfunctions [ J ]. Biochim Biophys Acta, 2012,1823:101-107.
  • 4翟妞,张业,沈珝琲.PIH1D1对染色质重塑复合物SNF5亚基降解的影响[J].中国医学科学院学报,2009,31(6):756-759. 被引量:1
  • 5von Morgen P, Horejsi Z, Macurek L. Substrate recognitionand function of the R2TP complex in response to cellularstress [ J ]. Front Genet, 2015,6: 69.
  • 6Yoshida M, Saeki M, Egusa H, et al, RPAP3 splicing va-riant isoform 1 interacts with PIH1D1 to compose R2TPcomplex for cell survival [ J ]. Biochem Biophys Res Com-mun, 2013,430: 320-324.
  • 7Boulon S, Pradet-Balade B, Verheggen C, et al. HSP90and its R2TP/Prefoldin-like cochaperone are involved in thecytoplasmic assembly of RNA polymerase II [ J ] . Mol Cell,2010,39: 912-924.
  • 8Inoue M, Saeki M, Egusa H,et al. PIH1D1,a subunit ofR2TP complex,inhibits doxorubicin-induced apoptosis[ J].Biochem Biophys Res Commun, 2010, 403 : 340-344.

二级参考文献10

  • 1Zhao R,Kakihara Y,Gribun A,et al.Molecular chaper-one Hsp90 stabilizes Pih1/Nop17 to maintain R2TP complexactivity that regulates snoRNAaccumulation[].The Journal of Cell Biology.2008
  • 2Sohn DH,Lee KY,Lee CJ,et al.SRG3 interacts directlywith the major components of the SWI/SNF chromatin re-modeling complex and protects themfromproteasomal degra-dation[].Journal of Biological Chemistry.2007
  • 3McKenna ES,Sansam CJ,Cho YJ,et al.Loss of the epi-genetic tumor suppressor SNF5 leads to cancer without ge-nomic instability[].Molecular and Cellular Biology.2008
  • 4Gresh L,Bourachot T,Reimann T,et al.The SWI/SNFchromatin-remodeling complex subunit SNF5 is essential forhepatocyte differentiation[].EMBO Journal.2005
  • 5Gonzales FA,Zanchin AT,Lus JS,et al.Characterizationof Saccharomyces cerevisiae Nop17p,a novel Nop58p-inter-acting protein that is involved in Pre-rRNA processing[].Journal of Molecular Biology.2005
  • 6Sudarsanam,P.,Winston,F.The Swi/Snf family nucleosome-remodeling complexes and transcriptional control[].Trends in Genetics.2000
  • 7Roberts,CW,Galusha,SA,McMenamin,ME,Fletcher,CD,Orkin,SH.Haploinsufficiency of Snf5 (integrase interactor 1) predisposes to malignant rhabdoid tumors in mice[].Proceedings of the National Academy of Sciences of the United States of America.2000
  • 8Tsikitis M,Zhang Z,Edelman W,et al.Genetic ablation of Cyclin D1abrogates genesis of rhabdoid tumors resulting from Ini1loss[].Proceedings of the National Academy of Sciences of the United States of America.2005
  • 9Klochendler-Yeivin,A,Picarsky,E,Yaniv,M.Increased DNA damage sensitivity and apoptosis in cells lacking the Snf5/Ini1 subunit of the SWI/SNF chromatin remodeling complex[].Molecular and Cellular Biology.2006
  • 10Isakoff,MS,Sansam,CG,Tamayo,P,Subramanian,A,Evans,JA,Fillmore,CM,Wang,X,Biegel,JA,Pomeroy,SL,Mesirov,JP,Roberts,CW.Inactivation of the Snf5 tumor suppressor stimulates cell cycle progression and cooperates with p53 loss in oncogenic transformation[].Proceedings of the National Academy of Sciences of the United States of America.2005

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部