2000 Fiscal Year Final Research Report Summary
Cloning of APG2 and APG15
| Project/Area Number |
11680708
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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 | Teikyo University of Science and Technology |
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Principal Investigator |
OHSUMI Mariko Teikyo University of Science and Technology, Associate Professor, 理工学部, 教授 (40168927)
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| Project Period (FY) |
1999 – 2000
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| Keywords | S.cerevisiae / Autophagy / Cloning / APG Genes |
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| Research Abstract |
The APG15 gene of Saccharomyces cerevisiae was cloned from a yeast genomic library by complementation of sporulation-deficient phenotype ofapg15-1 mutant. Structural analysis of the obtained genomic fragment revealedthat the APG15 gene is YMR159c, APG16 gene. APG16 gene was obtained by two-hybrid screening with Apg12p as bait by Mizushima, et. al..
Null mutant allele for YMR159c (Δ ymr159c) was constructed and introduced in to yeast. All apg mutants including apg15 were crossed with Δ ymr159c cells , and every resulting diploid showed Apg+phenotypes, thus the YMR159c was named as APG16. Sequecing of apg15-1 mutant allele revealed that it is single nucleotide conversion (A to G), resulting a nonsence mutation at 83rd amino acid. We carefully re-examined the complememtation. Apg+ phenotype of tetrad from ang15-1/wild showed non-Mendelian segregation. From several lines of evidence we got the conclusion that the suppression of apg15-1 with Δ apg16 was not caused by APG16 gene but a suppressor in the background of the strain. Apg15-1 mutation is quite efficienly suppressive mutation by widely distributed suppressor.
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