1996 Fiscal Year Final Research Report Summary
CHARACTERIZATION OF SENESCENCE GENES IN HUMAN CELLS
| Project/Area Number |
06454675
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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 (1995-1996)
The University of Tokyo (1994) |
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Principal Investigator |
AYUSAWA Dai KIHARA INSTITTE FOR BIOLOGICAL RESEARCH DEPARTMENT OF BIOCHEMISTRY YOKOHAMA CITY UNIVERSITY PROFESSOR, 木原生物学研究所, 教授 (00142109)
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| Project Period (FY) |
1994 – 1996
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| Keywords | SENESCENCE / IMMORTALIZATION / GENE / CHROMOSOME 7 / GENE MAPPING / RADIATION HYBRID |
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| Research Abstract |
We have undertaken the following two projects in a given time, and obtained several new findings that contribute understanding of cellular senescence and imnrotalization in human cells.
1. Identification of a senescence / immortalizing gene on human chromosome 7
We made a panel of G418-resistant radiation hybrid cells form mouse A7 cells containing human chromosome 7 tagged with pSV2neo. By analysis of FISH,chromosome transfer, and Alu PCR,we selected radiation hybrids that induce senescence in immortal cell lines assigned to genetic colmnplemenation D and have a very limited amount of human DNA.The human DNA was mapped to chromosome 7 to molecularly clone the senescence gene in a YAC vector.
2. Characterization of a signal transduction pathway in cellular senescence
Inhibitors of cGMP-dependent protein kinases was found to block senescence induced by heat inactivation of SV40T antigen in SV40T-transformed immortal human fibroblasts. Using one of such inhibitors as a ligand for affinity chromatography followed by Western blot analysis, we have found that a protein phosphatase was induced and dephosphorylation of specific proteins occurred. The mechanism of induction of the phosphatase and the proteins dephosphorylated were examined in vino and in vitro using specific inhibitors of phosphatases and antibodies.
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