| Project/Area Number |
61480474
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
分子遺伝学・分子生理学
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| Research Institution | Kyoto University |
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Principal Investigator |
YANAGIBA Mitsuhiro Department of Biophysics,Faculty of Science,Kyoto University,Professor, 理学部, 教授 (80025428)
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| Project Period (FY) |
1986 – 1987
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| Project Status |
Completed (Fiscal Year 1987)
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| Budget Amount *help |
¥5,500,000 (Direct Cost: ¥5,500,000)
Fiscal Year 1987: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1986: ¥5,000,000 (Direct Cost: ¥5,000,000)
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| Keywords | cell division / mitosis / chromosome condensation / gene product / スピンドル装置 / 細胞周期 / 染色体 / スピンドル / 細胞核 / 分裂酵母 |
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| Research Abstract |
Temperature sensitive (ts)mutation nuc2-663 of the fission yeast S.pombe causes specifically the arrest of mitotic spindle movement. Resulting spindle is short and uniform in length but cannot elongate. Chromosomes condense so that mtaphase plate-like structure to which the spindle runs perpendicularly is formed. The nuc2 mutation appears to block the transition from mitotic metaphase to anaphase.In wild type or mutant cells at permissive temperature,the spindle elongates 5-times in length during anaphase.
nuc2^+ gene was cloned,and its nucleotide sequence together with RNase protection experiment predicted a 665-residues polypeptide (MW 76000) containing short repeats.Antiserum was raised againt polyptide produced in E. coli using the fused nuc2^+ gene.By Western blotting,a polypeptide (MW 67000) was detected in the extracts of S. pombe,of which the amount increased by introducing multi-copy plasmid containing nuc2^+ gene.By cellular fractionation and nuclear isolation,we found that p67 is predominatly present in the nuclear fraction,specifically in the 'nuclear-scaffold'fraction insoluble in 2M NaCl or 25mM LIS. Furthermore p^<76> apparently the precursor of p67 is accumulated in the mutant extracts as the soluble form.We speculate that p67 may be a component in the kinetochore complex or in the spindle apparatus essential for chromosome separation.
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