2000 Fiscal Year Final Research Report Summary
Molecular mechanism of the high incidence of cancer of Bloom's syndrome
| Project/Area Number |
11138207
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| Research Category |
Grant-in-Aid for Scientific Research on Priority Areas (A)
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| Allocation Type | Single-year Grants |
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| Research Institution | Tohoku University |
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Principal Investigator |
ENOMOTO Takemi Graduate school of Pharm.Sci.Tohoku University Professor, 大学院・薬学研究科, 教授 (80107383)
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| Co-Investigator(Kenkyū-buntansha) |
MASUKO Takashi Graduate school of Pharm.Sci.Tohoku University Associate Professor, 大学院・薬学研究科, 助教授 (30157200)
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| Project Period (FY) |
1999
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| Keywords | Bloom's syndrome / BLM / SCE / yeast SGS1 / SUMO-1 / targeted integration |
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| Research Abstract |
Bloom's syndrome (BS) is a rare genetic disorder and the cells from BS patients show genomic instability and an increased level of sister chromatid exchange (SCE). We analyzed the functions of yeast Sgs 1, which is the yeast homologue for BLM, as well as those of BLM in higher eukaryotic cells and obtained the following results.
1. We made mutated SGS1 genes coding a protein having equivalent missense mutations of BS patients and found that none of the mutated genes could suppress the higher sensitivity to MMS and HU and the elevated SCE of sgs 1 disruptants. Analyses of Sgs 1 functions indicated that the two different mechanisms are operated in Sgs 1 functions, one requiring DNA helicase activity and one not. In addition, genetic analyses indicated that Sgs 1 functions in the down stream of Mec 1 and the upstream of Rad 51.
2. We generated a monoclonal antibody specifically recognizing BLM protein and using this antibody, found that BLM is localized in speckled structures in the nucleus. We also obtained a piece of evidence indicating that BLM is associated with SUMO-1. Deletion experiments indicated that the amino acid region 238-586 of BLM is required for the localization in speckled structures.
3. We generated BLM^<-/-> and BLM^<-/->/RAD54^<-/-> DT40 cells from the chicken B lymphocyte line DT40 and found that SCE and targeted integration frequencies were increased remarkably in BLM^<-/-> cells. The results obtained with BLM^<-/->/RAD54^<-/-> cells indicated that a large portion of the SCE in BLM^<-/-> cells occurs via homologous recombination and more double strand breaks occur during DNA replication in the absence of BLM and some of these breaks are repaired by homologous recombination.
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[Publications] Siratori, M., Sakamoto, S., Enomoto, T., Sugimoto, M., Goto, M., Matsumoto, T., and Furuichi, Y.: "Detection by epitope-defined monoclonal antibodies of Werner DNA helicases in nucleoplasm and their upregulation by cell transformation and immortalization."J.Cell Biol.. 144. 1-9 (1999)
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「研究成果報告書概要(欧文)」より
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[Publications] Matsumoto, Y., Satoh-Ueno, K., Yoshimura, A., Hashimoto, Y., Enomoto, T., and Masuko, T.: "Identification and Immunological Characterization of a Novel 40-kDa Protein Linked to CD98 Antigen."Cell Struct.Funct.. 24. 217-226 (1999)
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[Publications] Shishido, T., Ohkawa, M., Itoh, A., Enomoto, T., Hashimoto, Y., and Masuko, T.: "Colocalization of GP125/CD98 with tropomyosin isoforms at the cell-cell adhesion boundary."J.Biochem.. (in press).
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「研究成果報告書概要(欧文)」より
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[Publications] Shishido, T., Uno, S., Kamohara, M., Tsuneoka-Suzuki, T., Hashimoto, Y., Enomoto, T., and Masuko, T.: "Transformation of Balb3T3 cells caused by overexpression of rat CD98 heavy chain (HC) requires its association with light chain : Missense mutation in a cysteine residue of CD98HC eliminates its transforming activity."Int.J.Cancer. (in press).
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「研究成果報告書概要(欧文)」より
