1997 Fiscal Year Final Research Report Summary
Study on initiation of DNA repication in cell-free system of Xenopus eggs
| Project/Area Number |
07680762
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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 |
Cell biology
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| Research Institution | Kumamoto University |
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Principal Investigator |
SHIODA Masaki Kumamoto University Fac.Science, Dept.Biological Science, Professor, 理学部, 教授 (80134526)
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| Project Period (FY) |
1995 – 1997
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| Keywords | Xenopus laevis / DNA rreplication / cell-free DNA replication / initiator protein / license factor / initation of DNA replication / MCM / DNA untwisting protein |
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| Research Abstract |
We purified a novel protein (designated to DUF) which has an activity to untwist DNA and open double strand DNA to single strand DNA from cell-free DNA synthetic system of Xenopus eggs. DUF was composed of 87 and 140 kDa polypeptides. Immunobloting exper iments using antibody against the 87 kDa polypeptide showed that behavior of DUF during a cell cycle or embryogenesis co-relates with DNA replication. Immunodepletion of DUF from the DNA synthetic system lead to inhibition of intiation of DNA replication. These results suggest that DUF participates in intiation on of DNA replication. We also surveeid protein factors which regulate intiation of DNA replication using antibody agaist proteins concerning to DNA replication (human P1Cdc46 and human P1). As results, we detected a protein complex composed of 5 prteeins containing MCM famliy which regulates intiaton of DNA replication, correspondiing too the license factor which allows DNA replication only once in a cell cycle. Further studies on the relationship between DUF and license factor are required to elucidate regulation of initiation of DNA replication.
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[Publications] Murakami-Murofushi, K., Kobayashi, S., Onimura, K., Matsumoto, M., Yoshida, S., Shoji, M., Murofushi, H.: "Selective inhibition of DNA polymerase-alpha family with chemically synthesized derivatives of PHYLPA,a unique Physarum lysophosphatidic acid." Biochim.Biophys.Acta. 1258. 57-60 (1995)
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[Publications] Takayanagi, S., Kawasaki, H., Sugimori, K., Yamada, T., Sugai, A., Ito, T., Yamasato, K., and Shioda, M.: "Sulfolobus hakonensis sp.nov., a novel species of acidothermophilic Archaeon." Int.J.System.Bacteriol. 46. 377-382 (1996)
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[Publications] Yokoyama, Y., Okubo, T., Ozawa, S., Nagai, F., Ushiyama, K., Kano, I., Shioda, M., Kubo, H., Takemura, M., Namiki, H., Yasugi, E., Oshima, M., Seyama, Y., and Kano, K.: "CPP32 activation during dolichyl phosphate-induced apoptosis in U937 leukemia cells." FEBS Lett.412. 153-156 (1997)
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[Publications] Yokoyama, Y., Ozawa, S., Seyama, Y., Namiki, H., Hatashi, Y., Kaji, K., Shirama, K., Shioda, M., and Kano, K.: "Enhancement of apoptosis in developing chick neural retina cells by basic fibroblast growth factor." J.Neurochem. 68. 2212-2215 (1997)
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