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2000 Fiscal Year Final Research Report Summary

Role of protein phosphorylation in Myulating RNA polymerase II transcription

Research Project

Project/Area Number 11680685
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Molecular biology
Research InstitutionSaitama Medical School

Principal Investigator

HISATAKE Koji  Saitama Medical School, Biochemistry, Associate Professor, 医学部, 助教授 (70271236)

Project Period (FY) 1999 – 2000
KeywordsGeneral Transcription Factor / Baculo virus / Protein Phosphorylation / Transcription / TFIIH / Capping Reaction
Research Abstract

1. Role of ERCC3 in transcriptional activation by GAL4-VP16
In basal transcription, ERCC3 helicase acts during promoter opening and promoter escape. We tested at which step ERCC3 helicase acts during transcriptional activation by GAL4-VP16. The effect of TFIIH mutants were first tested using premelted templates, and revealed that TFIIH functions during promoter escape in activated transcription. The effect is specific to activation. Using the various mutant templates, we found that activators act during a very early stage of transcription. The effect requires coactivator PC4. RNA polymerase II is unstable during the early phase of transcription and may abort transcription. With the use of ATP analogs, we found that RNA polymerase II may detach from the template when ERCC3 helicase activity is blocked. Taken together, transcriptional activation involves the enhancement of recruitment of transcriptional machinery and promoter escape. Rate constant of promtoer escape is not stimulated in activated transcription.
2. Role of TFIIH in transcription-coupled capping reactions
In collaborated with Dr.Mizumoto, we have developed a system in which transcription and capping reactions can occur together. The system includes recombinant factors except RNA polymerase II.In this system, capping reaction occurs simultaneously with transcription initiation. The molecular analyses of capping and its relation to transcription initiation are in progress.

  • Research Products

    (4 results)

All Other

All Publications (4 results)

  • [Publications] Ishiguro,A.: "The Rpb 6 subunit of fission yeast RNA polymerase II is a contact target of the transcription elongation factor TFIIS"Molecular and Cullular Biology. 20. 1263-70 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Wada,T.: "FACT relieves DSIF/NELF-mediated inhibition of transcriptional elongation and reveals functional differences between P-TEFb and TFIIH."Molecular Cell. 5. 1067-1072 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ishiguro, A: "The Rpb 6 subunit of fission yeart RNA polymerase II is a contant targect of the transcription elongation factor TFIIS"Molecular and Cellular Biology. 20. 1263-70 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Wada, T: "The Fact uliums DSIF/NELF mediated inhibition of transcriptional elongation and Miuals functional differences between p-TEF6 and TFIIH"Molecular Cell. 5. 1067-1072 (2000)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2002-03-26  

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