1999 Fiscal Year Final Research Report Summary
Cloning of an immortality-suppressor gene on human chromosome 7
| Project/Area Number |
09044234
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| Research Category |
Grant-in-Aid for Scientific Research (B).
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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 | Kihara Institute for Biological Research |
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Principal Investigator |
AYUSAWA Dai Kihara Institute for Biological Research Professor, 木原生物研究所, 教授 (00142109)
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| Co-Investigator(Kenkyū-buntansha) |
SUZUKI Toshikazu Kihara Institute for Biological Research Assistant Professor, 木原生物研究所, 助手 (70270527)
FUJII Michihiko Kihara Institute for Biological Research Assistant Professor, 木原生物研究所, 助手 (80285155)
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| Project Period (FY) |
1997 – 1999
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| Keywords | chromosome 7 / microcell-mediated chromosome transfer / YAC / BAC / radiation hybrid / senescence / telomerase / telomere / STS marker |
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| Research Abstract |
Approximately 50% of immortal human cell lines so far tested seem to have a mutation on a gene on human chromosome 7 since introduction of this chromosome by microcell-mediated chromosome transfer specifically suppress their division potential. To identify it, we prepared a panel of radiation hybrids which contain a small segment of human chromosome 7 under mouse cell background. By chromosome transfer and PCR with STS markers, we narrowed down the gene within a 2-3 Mb region of the chromosome. We planned to rescue human DNA form the hybrids with a newly developed TAR (Transformation Associated Recombination) cloning system in yeast. In collaboration with NIH that invented the cloning system, we prepared a YAC/BAC library from one radiation hybrid. These BAC clones were introduced to human Met5A cell line to examine their anti-proliferative activity against this cell line.
As by-products of this study, we have shown that the immortality suppresser gene negatively regulates the telomere maintenance mechanisms in particular human cell lines. Upon introduction of the gene into the telomerase-positive lines, the telomerase catalytic subunit gene was down-regulated before entering senescence and with shortening of telomeres. In the negative lines, their usually long telomeres were immediately shortened.
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[Publications] Fujii, M., Ide, A., Nakabayashi, K., Joguchi, A., Ogino, H., and Ayusawa, D.: "The introduction of dominant-negative p53 mutants suppresses temperature shift-induced senescence in immortal human fibroblasts expressing a thermolabile SV40 large T antigen."J. Biochem.. 125. 531-536 (1999)
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「研究成果報告書概要(欧文)」より
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[Publications] Michishita, E., Nakabayashi, K., Suzuki, T., Kaul, S. C., Ogino, H., Fujii, M., Mitsui, Y., and Ayusawa, D.: "5-Bromodeoxyuridine Induces Senescence-like Phenomena in Mammalian Cells Regardless of Cell Types or Species"J. Biochem.. 126. 1052-14059 (1999)
Description
「研究成果報告書概要(欧文)」より
