• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

Involvement of mRNA capping enzyme in transcription initiation

Research Project

Project/Area Number 14580630
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Structural biochemistry
Research InstitutionKitasato University

Principal Investigator

SHIBAGAKI Yoshio  Kitasato University, School of pharmaceutical Sciences, Lecturer, 薬学部, 講師 (90235565)

Co-Investigator(Kenkyū-buntansha) HISATAKE Koji  Saitama Medical School, Assistant Professor, 医学部, 助教授 (70271236)
MIZUMOTO Kiyohisa  Kitasato University, School of pharmaceutical Sciences, Professor, 薬学部, 教授 (80092344)
Project Period (FY) 2002 – 2004
Project Status Completed (Fiscal Year 2004)
Budget Amount *help
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2004: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2003: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2002: ¥1,800,000 (Direct Cost: ¥1,800,000)
KeywordsCapping enzyme / C-terminal domain / RNA polymerase II / in vitro transcription / Transcription factor / Protein phosphorylation
Research Abstract

The mRNA capping occurs soon after transcription initiation and before other processing events. The several mRNA processing factors are coupled to transcription through binding to the C-terminal domain of RNA polymerase II (CTD). mRNA-capping enzyme directly binds to the phosphorylated CTD, and catalyzes the co-transcriptional cap formation. However, the precise mechanism of the involvement of capping enzyme in pol II transcription has been unclear.
(1) To examine the involvement of capping enzymes in pol II transcription, we developed in vitro transcription/capping system using immobilized DNA template. Immobilized templates were incubated with HeLa nuclear extract to form initiation complex (PICs) on promoter region. After washing to remove unbound proteins, PICs were subjected to elongation reaction in the presence of 3'-O-methly CTP in order to stall elongation complex at the first C residue. Using this system, we found that capping occurs in 18-19 nt RNA length in the case of adenovirus major late promoter.
(2) To investigate the functional interaction of capping enzyme and pol II, we subjected highly purified calf thymus pol II to capping assay with various kinds of CTD kinases. Capped RNAs were separated by Boronate-affinity PAGE (BAGE) to evaluate capping efficiency. We found that non-phosphorylated pol II stimulated capping activity about 2.5-fold. Furthermore, CAK phosphorylated pol II increased in capping activity about 1.8-fold compared with non-phosphorylated pol II. These results suggest that the CAK phosphorylation of pol II CTD is important for activation of capping enzyme and other portion of pol II also may involve in this activation.

Report

(4 results)
  • 2004 Annual Research Report   Final Research Report Summary
  • 2003 Annual Research Report
  • 2002 Annual Research Report
  • Research Products

    (21 results)

All 2004 2003 2002 Other

All Journal Article (13 results) Publications (8 results)

  • [Journal Article] Functional interaction of mRNA capping enzyme with RNA polymerase II2004

    • Author(s)
      和田京子
    • Journal Title

      生化学 76・8

      Pages: 855-855

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Annual Research Report 2004 Final Research Report Summary
  • [Journal Article] Functional interaction of mRNA capping enzyme with RNA polymerase II2004

    • Author(s)
      Wada, K., Shibagaki, Y., Hisatake, K., Fukuda, A., Mizumoto, K.
    • Journal Title

      Seikagaku 76

      Pages: 855-855

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] リン酸化RNAポリメラーゼII結合蛋白質の機能解析2004

    • Author(s)
      広瀬 豊
    • Journal Title

      第6回RNAミーティング要旨集

      Pages: 16-16

    • Related Report
      2004 Annual Research Report
  • [Journal Article] mRNAキャッピング酵素とRNA polymerase IIの相互作用2004

    • Author(s)
      和田京子
    • Journal Title

      第6回RNAミーティング要旨集

      Pages: 118-118

    • Related Report
      2004 Annual Research Report
  • [Journal Article] RNA polymerase II 転写初期反応とキャッピングとの相関2004

    • Author(s)
      小松哲郎
    • Journal Title

      第26回日本分子生物学会年会講演要旨集

      Pages: 471-471

    • Related Report
      2004 Annual Research Report
  • [Journal Article] mRNAキャッピング酵素活性に対するRNA polymerase II CTDのリン酸化の影響2004

    • Author(s)
      和田京子
    • Journal Title

      第26回日本分子生物学会年会講演要旨集

      Pages: 471-471

    • Related Report
      2004 Annual Research Report
  • [Journal Article] リン酸化RNAポリメラーゼII結合蛋白質の機能解析2004

    • Author(s)
      広瀬 豊
    • Journal Title

      第26回日本分子生物学会年会講演要旨集

      Pages: 471-471

    • Related Report
      2004 Annual Research Report
  • [Journal Article] 転写初期反応におけるmRNAキャッピング酵素の役割2003

    • Author(s)
      柴垣芳夫
    • Journal Title

      生化学

      Pages: 939-939

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] 転写初期反応におけるRNA polymerase IIによるmRNAキャッピング化酵素の役割2003

    • Author(s)
      柴垣芳夫
    • Journal Title

      国際シンポジウムRNAフロンティア

      Pages: 175-175

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] The role of mRNA capping enzyme in pol II transcription initiation2003

    • Author(s)
      Shibagaki, Y., Nojima, T., Hisatake, K., Fukuda, A., Mizumoto, K.
    • Journal Title

      Seikagaku 75

      Pages: 939-939

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] The role of mRNA capping enzyme in pol II transcription initiation2003

    • Author(s)
      Shibagaki, Y., Nojima, T., Matsui, H., Hisatake, K., Fukuda, A., Mizumoto, K.
    • Journal Title

      The New Frontier of RNA Science RNA 2003 Kyoto Abstract

      Pages: 175-175

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] RNA polymerase II転写複合体へのキャッピング酵素の組み込み2002

    • Author(s)
      柴垣芳夫
    • Journal Title

      生化学

      Pages: 1001-1001

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Involvement of mRNA capping enzyme in pol II initiation complex2002

    • Author(s)
      Shibagaki, Y., Hisatake, K., Fukuda, A., Fukamachi, N., Tsukamoto, T., Mizumoto, K.
    • Journal Title

      Seikagaku 74

      Pages: 1001-1001

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Publications] 柴垣芳夫: "Involvement of mRNA capping enzyme in transcription initiation"生化学. 75. 939 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] 松井博紀: "RNA polymerase II転写初期反応におけるmRNAキャッピング酵素の役割"第26回日本分子生物学会年会講演要旨集. 851 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] 和田京子: "mRNAキャッピング酵素とRNA polymerase IIの相互作用"第26回日本分子生物学会年会講演要旨集. 852 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] 久武幸司: "TFIIHのヘリカー背とキナーゼ活性の機能解析"第26回日本分子生物学会年会講演要旨集. 391 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Yoshio Shibagaki: "Involvement of mRNA capping enzyme in transcription initiation"Abstracts of RNA 2003 KYOTO "The New Frontier of RNA Science". 175 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] 柴垣芳夫: "RNAポリメラーゼII転写開始複合体へのキャッピング酵素の組み込み"生化学. 74. 1001 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] 柴垣芳夫: "転写初期反応におけるmRNAキャッピング酵素の役割"第25回日本分子生物学会年会講演要旨集. 527 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] 野島孝之: "転写開始複合体へのmRNAキャッピング酵素の組み込み"第25回日本分子生物学会年会講演要旨集. 527 (2003)

    • Related Report
      2002 Annual Research Report

URL: 

Published: 2002-04-01   Modified: 2016-04-21  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi