1993 Fiscal Year Final Research Report Summary
STUDY OF NEW INOSITOL-MEDIATED SIGNAL TRANSDUCTION MECHANISMS ON GENE CLANING
| Project/Area Number |
04833021
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
分子細胞生物学
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| Research Institution | KYUSYU INSTITUTE OF TECHNOLOGY |
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Principal Investigator |
NIKAWA Jun-ichi KYUSYU INSTITUTE OF TECHNOLOGY, FACULTY OF COMPUTER SCIENCE AND SYSTEMS ENGINEERING, ASSOCIATE PROFESSOR, 情報工学部, 助教授 (00134271)
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| Project Period (FY) |
1992 – 1993
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| Keywords | inositol / transcriptional regulation / gene cloning / promoter / yeast |
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| Research Abstract |
The octamer sequence TTCACATG is required for expression and regulation of inositol/choline-regulated genes of Saccharomyces cerevisiae, such as ITR1 which encodes inositol transporter I.In theis study, we constructed the yeast strain of which HIS3 expression is controlled under the ITR1 promoter. Using this strain, we isolated three genes that, when introduced in multicopies, alleviate the repression caused by inositol via ITR1 promoter. Morthern blot analysis the revealed that two of these three genes, designated as DIE1 and DIE2, clearly increased the expression of ITR1. DIE2 is more effective on itr1 expression than DIE1. Gene disruption experiments revealed that DIE1 was essential for the expression of ITR1 but DIE2 was not. The sequence of DIE1 gene shows that it is indentical to INO2 (also celled SCS1) which encodes a pretein required for the expression of INO1, which encode inositol 1- phosphatase synthase. DIE2 is a new gene and is capable of encoding 525 amino acid residues with a calculated molecular weight if 61, 789. Experiments using lacZ fusion genes showed that multicopy DIE2 resulted the increase of the expression of ITR1 and INO1. These results strongly suggest that DIE2 gene product may have an important regulatory function for not only ITR1 gene expression but slso INO1 gene expression.
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