Mechanism of transcription of ribosomal DNA in vitro
| Project/Area Number |
10680656
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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 | Saitama Medical School |
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Principal Investigator |
NOGI Yasuhisa Department of Medicine, Saitama Medical School, Professor, 医学部, 教授 (60101937)
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| Project Period (FY) |
1998 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 1999: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1998: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Keywords | the fission yeast / in vitro / RNA polymerase I / 分裂酵素 / rDNA |
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| Research Abstract |
1. SpRPA12 subunit is required for efficient transcription of ribosomal DNA (rDNA) in the fission yeast in vitro. (1) Analysis of function of SpRPA1 2 subunit in vitro. S100 fraction was prepared from the wild-type and Sprpa12 disruptant strains to compare the rDNA transcription activity of the mutant relative to wild-type. In vitro transcription was carried out using the respective extracts and the rRNA synthesized in vitro was detected by Si nuclease mapping. The results have shown that Sprpa12 mutant extract retained only 10% transcription activity compared to wild-type, indicating that SpRPA12 subunit is required for the efficient transcription of rDNA in the fission yeast. The thermolability of the transcription activity of the respective extracts was also examined, suggesting that SpRPA12 is required for the thermostability of the RNA polymerase I or the presumed preinitiation complex.. (2) Analysis of function of SpRPA12 in vivo. The deletion analysis of the C-terminal region of SpRPA12 was performed and the results have shown that the C-terminal zinc-finger motif that is conserved among RPA12 homologs is not required for the function of SpRPA12. The deletion analysis of the N-terminal region containing the N-terminal zinc-finger motif is now on progress.
2. Characterization of RPA42 subunit. We have isolated and characterized RPA42 subunit of the fission yeast Pol I. The results have shown that RPA42 interacts specifically with RPA17 subunit and both of them show homologies to alpha subunit of RNA polymerase in prokaryotes, suggesting that RPA42 and RPA17 form a core of Pol I.
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Report
(3 results)
Research Products
(13 results)
