2003 Fiscal Year Final Research Report Summary
Study on the regulation of DNA replication by the modulation of chromatin
| Project/Area Number |
14580692
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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 | Yamaguchi University (2003)
The University of Tokyo (2002) |
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Principal Investigator |
MUROFUSHI Hiromu Yamaguchi University, Faculty of Science, Professor, 理学部, 教授 (70101128)
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| Project Period (FY) |
2002 – 2003
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| Keywords | Xenopus egg / DNA replication / Cdc68p / Spt16p / SSRP1 / DNA repair / RPA / telomerase / DNA helicase |
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| Research Abstract |
1. Alteration of chromatin structure by DUF
Chromatin was reconstituted in Xenopus egg extracts deprived of DUF with the aid of anti-DUF antibody and nuclease sensitivity of the chromatin was measured. It was revealed that chromatin made in the absence of DUF was less sensitive to nuclease than control chromatin made in the presence of DUF. It was also shown that DUF interacts not only with duplex DNA but also with core histones. These data suggest that DUF makes more relaxed chromatin accessible for replication factors
2. Search for factors interacting with DUF
Factors interacting with 140 kDa subunit of human DUF were screened by the yeast two-hybrid system. It was shown that CLKI, GU2, ribosomal proteins L15, and S26 interacted with DUF. Co-immunoprecipitation experiments revealed that Orc1 binds to DUF, suggesting that DUF in complex with ORC is involved in the initiation of DNA replication
3. Dynamics of RPA in DNA repair
Dynamics of RPA was investigated in Xenopus egg cell-free system. Double strand breaks in DNA induced RPA foci in the sites where DNA repair synthesis occurred. Inhibition of ATM/ATR showed little effects on the formation of RPA foci. These data suggest that DNA damage checkpoint mechanism is not involved in the formation of RPA foci (in collaboration with T.Enomoto, Tohoku University)
4. Study on DNA helicase with affinity for telomerase
In order to find new factors involved in DNA replication of mammalian telomeres, proteins co-purified with telomerase were investigated. cDNA cloning revealed that a protein with motifs of DNA helicase was co-purified with telomerase. (in collaboration with K.Murakami-Murofushi, Ochanomizu University). Expression of the protein was performed using the baculovirus system
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Research Products
(13 results)
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[Publications] Seo, H., Okuhara, K., Kurumizaka, H., Yamada, T., Shibata, T., Ohta, K., Akiyama, T., Murofushi, H.: "Incorporation of DUF/FACT into chromatin enhances the accessibility of nucleosomal DNA"Biochem.Biophys.Res.Commun.. 303. 8-13 (2003)
Description
「研究成果報告書概要(欧文)」より
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[Publications] Kunimoto, S., Murofushi, W., Yamatsu, L, Hasegawa, Y., Sasaki, N., Kobayashi, S., Kobayashi, T., Murofushi, H., Murakami-Murofushi, K.: "Cholesteryl glucoside-induced protection against gastric ulcer"Cell Structe.Funct.. 28. 179-186 (2003)
Description
「研究成果報告書概要(欧文)」より
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[Publications] Kobayashi, T., Tada, S., Tsuyama, T., Murofushi, H., Seki, M., Enomoto, T.: "Focus-formation of replication protein A, activation of checkpoint system and DNA repair synthesis induced by DNA double-strand breaks in Xenopus egg extract"J.Cell Sci.. 115. 3159-3169 (2002)
Description
「研究成果報告書概要(欧文)」より
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[Publications] Kobayashi, T., Tada, S., Tsuyama, T., Murofushi, H., Seki, M., Enomoto, T.: "Effect of topoisomerase inhibitors on the formation of replication protein A-foci in nuclei reconstituted in Xenopus egg extract"J.Health Sci.. 48. 296-301 (2002)
Description
「研究成果報告書概要(欧文)」より
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[Publications] Iida, J., Itoh, T.J., Hotani, H., Nishiyama, K., Murofushi, H., Bulinski, J.C., Hisanaga, S.: "The projection domain of MAP4 suppresses the microtubule-bundling activity of the microtubule-binding domain"J.Mol.Biol.. 320. 97-106 (2002)
Description
「研究成果報告書概要(欧文)」より
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[Publications] Kunimoto, S., Murofushi, W., Kai, H., Ishida, Y., Uchiyama, A., Kobayashi, T., Kobayashi, S., Murofushi, H., Murakami-Murofushi, K.: "Steryl glucoside is a lipid mediator in stress-responsive signal transduction"Cell Struct.Funct.. 27. 157-162 (2002)
Description
「研究成果報告書概要(欧文)」より
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[Publications] Murakami-Murofushi, K., Uchiyama, A., Fujiwara, Y., Kobayashi, T., Kobayashi, S., Mukai, M, Murofushi, H., Tigyi, G.: "Biological functions of navel lipid mediator, cyclic phoshatidic acid"Biochim.Biophys.Acta. 1582. 1-7 (2002)
Description
「研究成果報告書概要(欧文)」より
