Functional characterization of phosphorylated RNA polyrnerase II interacting factors
| Project/Area Number |
14580682
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Molecular biology
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| Research Institution | KANAZAWA UNIVERSITY |
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Principal Investigator |
HIROSE Yutaka KANAZAWA UNIVERSITY, Canoer Research Institute, Assistant Professor, がん研究所, 助手 (00218851)
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| Project Period (FY) |
2002 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 2003: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2002: ¥1,700,000 (Direct Cost: ¥1,700,000)
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| Keywords | gene expression / transcription / mRNA processing / RNA polymerase II / phosphorylation / WW domain / gene knock-out |
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| Research Abstract |
Phosphorylation of the carboxy-terminal domain (CTD) of RNA polymerase II largest subunit has important roles both in transcription and in coupling transcription to pre-mRNA processing. To better understand the molecular mechanism in which transcription coordinates with pre-mRNA processing, I have identified and characterized human factors that can directly interact with the phosphorylated CTD (pCTD).Recently I reported a novel human protein PCIF1 as a pCTD interacting factor. To investigate cellular functions of PCIF1, I disrupted the PCIF1 gene in the chicken B-cell line DT4O. I have established two independent mutant DT40 cell lines in which all three copies of PCIF1 gene are disrupted by homologous recombination. One clone is a homozygous PCIF1-null mutant and the other is a conditional knock out cell line in which PCIF1 protein is expressed from a chicken PCIF1 cDNA under control of a tetracycline-repressible promoter.Addition of doxycyclineto the cells results in depletion of PCIF1 protein within several days but not in growth defect. Thus, chicken PCIF1 is not essential for cell growth. Depletion of PC1FI in DT40 cells did not significantly affect neither the phosphorylation status of the CTD nor the methylation status of the histone H3 N-terminus region.PCIF1-deficient cells exhibited normal heat shock-response, as measured by inducible expression of heat shock genes.
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Report
(3 results)
Research Products
(5 results)
