| Project/Area Number |
04404025
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| Research Category |
Grant-in-Aid for General Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Research Field |
General medical chemistry
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| Research Institution | Saitama Medical School |
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Principal Investigator |
MURAMATSU Masami Saitama Medical School Fac.of Med.Professor, 医学部, 教授 (10035454)
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| Co-Investigator(Kenkyū-buntansha) |
MAKINO Yasutaka Chiba University Fac.of Science Assistant, 理学部, 助手 (20240989)
TAMURA Taka-aki Chiba University Fac.of Science Professor, 理学部, 教授 (30112692)
SUZUKI Toshiya Saitama Medical School Fac.of Med.Assistant, 医学部, 助手 (90216416)
YAMAMOTO Kazuo Saitama Medical School Fac.of Med.Assistant, 医学部, 助手 (70255123)
NOGI Yasuhisa Saitama Medical School Fac.of Med.Assistant Professor, 医学部, 助教授 (60101937)
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| Project Period (FY) |
1992 – 1993
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| Project Status |
Completed (Fiscal Year 1993)
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| Budget Amount *help |
¥24,000,000 (Direct Cost: ¥24,000,000)
Fiscal Year 1993: ¥8,000,000 (Direct Cost: ¥8,000,000)
Fiscal Year 1992: ¥16,000,000 (Direct Cost: ¥16,000,000)
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| Keywords | rDNA / RNA polymerase / UBF / Transcriptional Regulation / FM3A / MH134 / RPA40 / PAF53 / RNAポリメラーゼI(PolI) / 転写因子 / PIN / ERCC3 / ペレット転写法 / RNAポリメラーゼI / 核小体 / HMG-box / NLS / 酸性ドメイン |
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| Research Abstract |
The major accomplishments during the three year period are summarized as follows.
1. Elucidation of the nucleolar targeting mechanism of mouse UBF (mUBF) and analysis of the transcription regulatory region of the mUBF gene.
By examining various domain deletion mutants of mUBF, it was found that the mUBF goes into the nucleus by NLS present in HMG box and accumulated into the nucleous by the strong affinity of the HMGbox1 and acidic domain to rDNA and SL1, respectively. On the other hand, by using upstream deletion mutants of mUBF connected to CAT gene, the mUBF gene has been found to have multiple SP1 binding sites working like one of the house keeping genes.
2. Identification of an RNA polymerase I inhibitory protein (PIN) in growth-arrested FM3A cells.
We have identified a protein which specifically inhibits rDNA transcription in vitro in extracts from mouse FM3A cells that have been deprived of serum and growth-arrested. This factor, designated PIN, appears to inhibit the start of pol I from the initiation complex.
3. Determination of subunit organization of mouse pol I and the cloning of RPA40 subunit and PAF 53 (polymerase associated factor 53).
We have developed a new purification procedure for RNA polymerase I (pol I) from mouse MH134 cells and analyzed the subunit composition. Unlike previous reports, mouse pol I was found to have about twelve small subunits in addition to two large subunits, corresponding in major part to those of yeast S.cerevisiae pol I.This has shown a strong conservation of pol I structure during evolution. Furthermore, a pol I associated factor with Mr=54KD was cloned and was found to stimulate specific rDNA transcription by interacting both with pol I and UBF.
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