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Analysis of gene regulatory function of TFIIF and elongin

Research Project

Project/Area Number 09470519
Research Category

Grant-in-Aid for Scientific Research (B).

Allocation TypeSingle-year Grants
Section一般
Research Field Human genetics
Research InstitutionTokyo Medical and Dental Univerisry

Principal Investigator

SHIGETAKA Kitajima  Medical Research Institute, Tokyo Medical and Dental Univerisry Professor, 難治疾患研究所, 教授 (30186241)

Co-Investigator(Kenkyū-buntansha) ATORU Oshiro  Medical Research Institue, Tokyo Medical and Dental Univerisry Assistant Professor, 難治疾患研究所, 助手 (30160485)
TERUMASA Tuchiya  Medical Research Insitute, Tokyo Medical and Dental Univerisry Assistant Professor, 難治疾患研究所, 助手 (20242109)
YUKIO Yasukochi  Medical Research Institue, Tokyo Medical and Dental Univerisry Professor, 難治疾患研究所, 教授 (60037398)
TEIJIRO Aso  Cancer Research Institue, Investigator, 癌研究会研究所, 研究員 (20291289)
Project Period (FY) 1997 – 2000
Project Status Completed (Fiscal Year 2000)
Budget Amount *help
¥13,300,000 (Direct Cost: ¥13,300,000)
Fiscal Year 2000: ¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1999: ¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1998: ¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 1997: ¥5,500,000 (Direct Cost: ¥5,500,000)
KeywordsTFIIF / Tripartite structure of RAP74 / HIV-Tat / Transcriptional elongation / Transcriptional recycling / Elongin / Testis-specific A2 / Gene targeting of elongin / RNAポリメラーゼII / 組織特異的因子 / 開始型 / 伸長型 / トリプシン分解 / 組織特異的エロンガン / 転写制御 / p32 / p30 / RNAポリメラーゼIIホロ酵素
Research Abstract

TFIIF, which recruits RNA polymerase II (Pol II) into preinitiation complex formed at gene promoter, can also function as an elongation factor for mRNA synthesis by Pol II, Elongin is a general elongation factor which may be involved in oncogenesis in VHL disease. To understand regu1atory mechanism of transcriptional elonagtion, we investigated function of TFIIF and elongin in this study.
1) TFIIF
RAP74 subunit of TFIIF is predicted to have tri-partite structure ; N- and C-terminal domains, and central region. We identified autoantibodies against RAP74 subunit of TFIIF in sera from autoimmune disease patients which selectively recognize the central portion of RAP74. By 2-hybrid screen, p32 was cloned as RAP74-interacting gene. p32 could bind to central region of RAP74 and HIV-Tat in vitro, and was capable for accelerating elongation by Pol II in the presence of HIV-Tat. p32 may be implicated in replicative growth of HIV.We could not clone FCP1, which binds to C-terminal of RAP74 and dephosphorylates CTD of Pol II by 2-hybrid screening. FCP1 was reported by Greenblatt et al. We expressed and purified recombinant TFIIF through co-infection of baculoviruses for RAP30 and 74. Using this, structural analysis of free, initiation, and elongation forms of TFIIF was performed. Tripartite structure of RAP74, its outer localization in TFIIF, and inner location of RAP30 was revealed. We are trying to crystallize TFIIF molecule for structural analysis.
2) Elongin
We cloned a novel form of elongin, A2, from EST on database. A2 is expressed preferentially in the testis while A is ubiquitously expressed. Recombinant A2 is capable for stimulating elongation of Pol II in vitro. A2 may function in expression of testis-specific genes. Furthermore, we identified A3, another novel form of elongin family. We are now characterizing its strucutre and function. Our gene knock-out project is under investigation, We screened ES cells which are heterologous for elongin A gene.

Report

(5 results)
  • 2000 Annual Research Report   Final Research Report Summary
  • 1999 Annual Research Report
  • 1998 Annual Research Report
  • 1997 Annual Research Report
  • Research Products

    (32 results)

All Other

All Publications (32 results)

  • [Publications] Cai Y,Kitajima S,Etoh F,Kinoshita S,Okubo K,Hamasaki N.: "Autoantibodies against human general transcription initiation factor TFIIF in sera of autoimmune diseases."Clin.Exp.Immunol.. 109. 488-494 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Muta T,Kang D,Kitajima S,Fujiwara T,Hamasaki N.: "p32 protein, a splicing factor 2-associated protein, is localized in mitochondrial matrix and functionally important in maintaining oxidative phosphorylation"J.Biol.Chem.. 272. 24363-24370 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kitajima S,Cai Y,Tanaka M,Nawa T,Oshiro S.: "Mechanism of transcription by RNA polymerase II- a molecular basis of regulated gene expression"J.Med.Dent.Sci.. 45. 59-67 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Cai Y,Mitsuyasu H,Oshiro S,Hamasaki N,Kitajima S.: "Structure of the human transcription factor TFIIF revealed by limited proteolysis with trypsin"FEBS Letters. 435. 191-194 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Aso T,Yamazaki K,Amimoto K,Higashi H,Kitajima S,Hatakeyama M.: "Identification and characterization of elongin A2, a new member of the elongin family of transcription factors, specifically expressed in the testis"J.Biol.Chem.. 275. 6546-6552 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Aso T,Yamazaki K,Aigaki T,Kitajima S.: "Drosophila von Hippel-Lindau tumor suppressor complex possesses E3 ubiquitin ligase activity"Biochem.Biophys.Res.Commun.. 276. 355-361 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Cai Y, Kitajima S, Etoh F, Kinoshita S, Okubo K, Hamasaki N.: "Autoantibodies against human general transcription initiation factor TFIIF in sera of autoimmune diseases."Clin.Exp.Immunol.. 109. 488-494 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Muta T, Kang D, Kitajima S, Fujiwara T, Hamasaki N.: "p32 protein, a splicing factor 2-associated protein, is localized in mitochondrial matrix and functionally important in maintaining oxidative phosphorylation."J.Biol.Chem. 272. 24363-24370 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kitajima S, Cai Y, Tanaka M, Nawa T, Oshiro S.: "Mechanism of transcription by RNA polymerase II-a molecular basis ofregulated gene expression-"J.Med.Dent.Sci.. 45. 59-67 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Cai Y, Mitsuyasu H, Zhang C, Oshiro S, Hamasaki N, Kitajima S.: "Structure of the human transcription factor TFIIF revealed by limited proteolysis with trypsin"FEBS Letters. 435. 191-194 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Aso T, Yamazaki K, Amimoto K, Higashi H, Kitajima S, Hatakeyama M.: "Identification and characterization of elongin A2, a new member of the elongin family of transcription factors, specifically expressed in the testis"J.Biol.Chem. 275. 6546-6552 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Aso T, Yamazaki K, Aigaki T, Kitajima S.: "Drosophila von Hippel-Lindau tumor supressor complex possesses E3 ubiquitin ligase activity"Biochem.Biophys.Res.Commun. 276. 355-361 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Aso,T., Yamazaki,K., Amimoto,K., Kuroiwa,A., Kitajima,S., et al: "Identification and characterization of elongin A2, a new member of the elongin family of transcription factor, specifically expressed in the testis"Journal of Biological Chemistry. 275. 6546-6552 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Cai,Y., Zhang,C., Nawa,T., Aso,T., Tanaka,M., Oshiro,S., Kitajima,S.: "Homocysteine-responsive ATF3 gene expression in human vascular endothelial cells : activation of c-Jun NH2-terminal kinase and promoter response element"Blood. 96. 2140-2148 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Nawa,T., Nawa,TM., Cai,Y., Zhang,C., Uchimura,I., Narumi,S., Kitajima,S.: "Repression of TNF-a-induced E-selectin expression by PPAR activators : involvement of transcriptional repressor ATF3/LRF1"Biochem Biophys.Res.Commun.. 275(2). 406-411 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Aso,T., Yamazaki,K., Aigaki,T., Kitajima,S.: "Drosophila von Hippel-Lindau tumor suppressor complex possesses E3 ubuquitin ligase activity"Biochem Biophys.Res.Commun.. 276. 355-361 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] 北嶋繁孝,麻生悌二郎,山崎勝久: "転写伸長因子の機能:TFIIFとエロンガン"蛋白質核酸酵素. 45. 1513-1522 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Verhoef, K. et al: "Evolution of the HIV-1 LTR promoter by conversion of an NF-KB enhancer element into a GABP binding site"J. Virology. 73. 1331-1340 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Hayakawa, H. et al: "Metabolic fate of oxidized guanine ribonucleotides in mammalian cells."Biochemistry. 38. 3610-3614 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Aso, T., et al: "Identification and characterization of elongin A2, a new member of the elongin family of transcription elongation factors, specifically expressed in tests"J. Biol. Chem.. (in press).

    • Related Report
      1999 Annual Research Report
  • [Publications] Nakamura,R.,et al: "The PC motif:a novel and erolutionally conserved sequence inrolued in interaction between p40 and p67 phox,SH3 domain-containing cytosolic factoy of NADPH cxidase" Eur.J.Biochem.251. 583-589 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Oshiro,S.,et al: "Aluminum taken up by transferrin-independent uptake affects the iron metabolism in rat cortical cells" J.Biochem.123. 42-46 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Kitajima,S.,et al: "Mechanism of transcinption by RNA polymerasell -a molecular basis of rasulated gene expression-" J.Medical & Dental Sciences. 45. 59-67 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Sugimoto,R.,et al: "Cloning and characterization of the Hakata antigen,a member of the ficolin/opsonin p35 lectin family" J.Biol.Chem.273. 20721-20727 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Cai,Y.,et al: "Structure of the human transinption factor TFIIF reuealed by limited proteolysis with trypsin" FEBS Letters. 435. 191-194 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Verhoef,K.,et al: "Eoolution of the HIV-1 LTR promoter by conversion of an NF-KB enhemcer element into a GABP binding sire" J.Virology. (in press). (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Kamura,T., et al: "Characterization of the human thrombopoieting gene promuter : A possible role of an Ets-transciption foctor E4TF1/GABP" J.Biol.Chem.272. 11361-11368 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Chibazakura,T., et al: "Phosphoyiation of human geneol tronsciption factons TATA-binding protein and IIB by DNA-dependent protein kinare : Synengistic stomelation of RNA poiymeooeII besal trowonppon in vitro" Eur.J.Biochem.247. 1166-1173 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Cai,Y., et al: "Autoantibodies agcuist human general tromeciption initiation factor TFIIF in sera of autoimmune diseases" Clin.Exp.Immunol.109(3). 488-494 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Muta,T., et al: "p32 protein,a splicing foctor 2-associbted protein,is localized in mitochosduial matrix and funetionelly important in mantainirg oxidetive phosphoryletion" J.Biol.Chem.272. 24363-24370 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Aso,T., et al: "Molecular cloning of DNAs encoding the reguletory subunits of Elongin from Sacchoromyces cerevisiae and Drosophila melanogsoter" Biochem.Biophys.Res.Commun.241. 334-340 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Pan,G., et al: "Interaction of elongation factors TFIIS and elongin A uidh a human RNA poiymerae II holoenjyne capable of protein-specific initiation and responave to trorsuiptind activators" J.Biol.Chem.272. 24563-24571 (1997)

    • Related Report
      1997 Annual Research Report

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Published: 1997-04-01   Modified: 2016-04-21  

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