| Project/Area Number |
02808030
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
物質生物化学
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| Research Institution | Kitasato University School of Hygienic Science |
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Principal Investigator |
OHTSUKI Kenzo Kitasato Univ. School of Hygienic Science, Assistant Professor, 衛生学部, 助教授 (60124559)
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| Co-Investigator(Kenkyū-buntansha) |
OH-ISHI Masamichi Kitasato Univ. School of Hygienic Science, Associate Professor, 衛生学部, 助手 (40233027)
高尾 雅 北里大学, 衛生学部, 助手 (70216612)
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| Project Period (FY) |
1990 – 1991
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| Project Status |
Completed (Fiscal Year 1991)
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| Budget Amount *help |
¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1991: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1990: ¥1,000,000 (Direct Cost: ¥1,000,000)
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| Keywords | D-kinase / Casein kinase II / p98 DNA binding protein / Protein phosphorylation / Embryogenesis / Initiction of DNA relication / Differentiation / Proliferation / 特異活性化因子 / 特異リン酸化ポリペプチド / リン酸化ポリペプチドの生理活性 / 転写調節 |
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| Research Abstract |
I. Characterization of p98 as an effective phosllhate accellor for D-kinase
DNA-dependent protein kinase(D-kinase)had been purified to homogeneity from the 1.5M KCI extract of the unfertilized eggs of sea urchin. Using the purified kinase, a specific protein with a DNA binding ability was highly purified from the egg extract(or cell extract from malignant cells)as an effective phosphate acceptor for the kinase by means of phospho-cellulose, gel filtration on Sephacryl S300, heparin-affinity and DNA-affinity column chromatographies, successively. The biochemical characterization of the DNA binding protein revealed that(i)the protein(pI 9.9)was composed of two distinct subunits (alpha and beta)and was pecifically phosphorylated by D-kinase, which had a property similar to casein kinase 11(CK-II) ; (ii)sperm-protamine extremely stim'ulated its phosphorylation by the kinase in vitro.
II. The physiological role of p98
During the purification of p98 from the eggs of sea urchin and mouse maliganat cells, it was found that the protein was partially purified from the cell extracts as a complex with p53 and D-kinase. The experimental observations the(i)p53 could be cross-reacted with anti-serum of suppressor onoco-gene product(p53)and(ii)no suppressing activity of p53 after its phosphorylation by the kinase against the DNA replication in vitro was detected, suggest that the physiological activity p98 may be controlled by the p53 through its specific phosphorylation by the kinase in the presence of some activators, such as protamine and histone.
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