1997 Fiscal Year Final Research Report Summary
Regulation of Budding Timing by clock-related gene GTS1 in Saccharomyces cerevisiae.
| Project/Area Number |
07680766
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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 | Yamanashi Medical University |
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Principal Investigator |
MITSUI Kazuhiro Yamanashi Medical University Medicine Associate professor, 医学部, 助教授 (20174063)
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| Project Period (FY) |
1995 – 1997
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| Keywords | Saccharomyces cerevisiae / Biological rhythm / ABC protein / Glycolysis / Ultradian rhythm |
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| Research Abstract |
We isolated the GTS1 gene from Saccharomyces cerevisiae using per gene, which is clock gene of Drosophila, as probe. The GTS1 gene affected the timing of budding and cellsize of Saccharomyces cerevisiae. The GTS1 gene also affected the heat tolerance, sporulation, life span of S.cerevisiae. We searched the functional region of the gene product (Gtslp) using yeast Two-Hybrid system. We found that the Gtslp forms the homodimer in yeast cell via 18 amino acids region in carboxyl terminal half. Furthermore, we showed that the putative binding site of Gtslp is highly homologous to a portion of the cytoplasmic domain of some kind of multidrug resistance proteins and that Gtslp bound to the cytoplasmic domain of the multidrug resistance proteins in Two-Hybrid system. The GTS1 overexpressed strain was highly sensitive to Cadmium, Cyclohexymide compare to wildtype strain.
We got some clones that interact with Gtslp in vivo using Two-Hybrid system. Two of the clone were Glycolytic enzymes, Glyceraldehyde-3-phosphate dehydrogenase and Pyruvate kinase. We examined the effect of GTS1 on the synchronization of ultradian oscillation of the concentration of glycolytic metabolite NADH and showed that the frequency and amplitude of the oscillation were modulated by GTS1 in gene dosage manner.
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