1999 Fiscal Year Final Research Report Summary
ESTABLISHMENT OF HIGH-THROUGHPUT SCREENING FOR IMMUNOSUPPRESSANTS USING YEAST MODEL SYSTEM
| Project/Area Number |
09557009
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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 | 展開研究 |
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| Research Field |
General pharmacology
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| Research Institution | Kobe University |
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Principal Investigator |
KUNO Takayoshi KOBE UNIVERSITY, SCHOOL OF MEDICINE, DEPARTMENT OF PHARMACOLOGY, PROFESSOR, 医学部, 教授 (50144564)
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| Co-Investigator(Kenkyū-buntansha) |
TAMURA Kouichi FUJISAWA PHARMACEUTICAL CO., LTD., HEAD INVESTIGATOR, 開発研究所, 主任研究員
SHUNTOH Hisato KOBE UNIVERSITY, FACULTY OF HEALTH SCIENCES, ASSOCIATE PROFESSOR, 医学部, 助教授 (70206259)
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| Project Period (FY) |
1997 – 1999
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| Keywords | immunosuprressant / protein phosphorylation / protein phosphatase / screening / fission yeast / MAP kinase |
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| Research Abstract |
Isolation of fission yeast mutants that requires calcineurin phosphatse activity for their growth and their application for a high-throughput screening of immunosuppressants. When calcineurin gene was disrupted or FK506 (taclolimus) was added to the media, wild-type fission yeast cells were viable. We mutagenized wild-type fission yeast cells and found several immunosuppressant-sensitive mutants. It is indicated that these mutants are synthetically lethal with calcineurin disruption and the mutated genes are sharing an essential function with calcineurin. Several immunosuppressant-sensitive mutants also showed sensitivity to high temperature, suggesting those mutated genes are essential, and named immunosuppressant- and temperature-sensitive mutants (its mutants). We isolated 8 its mutants and identified mutated genes. All of these mutants required calcineurin activity for their viability. The screening procedure uses wild-type and several its mutant strains. If candidate drug inhibits the growth of its mutants but not of wild-type cells, it is indicated that the drug specifically inhibits calcineurin activity and should be a good candidate for a immunosuppressant. We also developed a novel screening system for an inhibitor of MAP kinase pathway using calcineurin disrupted fission yeast strain. These screening systems are readily available for both academic and industrial use.
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