1997 Fiscal Year Final Research Report Summary
Construction of analytical system for cardiac function of the phospholamban knock-out mouse
| Project/Area Number |
08557049
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
Circulatory organs internal medicine
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| Research Institution | Osaka University |
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Principal Investigator |
TADA Michihiko Osaka Univ., Medical School., Professor, 医学部, 教授 (90093434)
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| Co-Investigator(Kenkyū-buntansha) |
NISHIDA Masashi Osaka Univ., Medical School., Associate Prof., 医学部, 助手 (40283783)
HOSHIDA Shiro Osaka Univ., Medical School., Associate Prof., 医学部, 助手 (80238732)
KUZUYA Tsunehiko Osaka Univ., Medical School., Associate Prof., 医学部, 助教授 (80150340)
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| Project Period (FY) |
1996 – 1997
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| Keywords | phospholamban / cardiac myocytes / knock-out mouse / gene regulation / NF-YA / NF-YB |
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| Research Abstract |
We have been studying the properties of phospholamban (PLN) in regard to its physiological aspects. To understand its role in the intact heart, we attempted to create the PLN-knock out mouse. In the first step, we had to study the transcriptional regulation of PLN because of its unique expression in the cardiac myocytes. We analyzed a 5'-upstream region of the phospholamban gene. Using a series of deletion constructs, we demonstrated that the region from -96 to-78 base pairs containing the CCAAT motif is essential for transcription of this gene. This region specifically binds to nuclear proteins extracted from rat hearts, and gel shift assays using competitive oligonucleotides, antibodies and recombinant proteins show that this region binds to the NF-YA and NF-YB among the CCAAT motif-binding proteins. This region-dependent transcription in cells transfected with antisense cDNAs encoding NF-YA and NF-YB was decreased to approximately 50% of that seen in cells transfected with the same sense cDNAs. We therefore conclude that the region spanning -96 to -78 base pairs plays a critical role in expression of the phospholamban gene which is regulated by binding of the nuclear protein NF-Y.In the second step, we started creating PLN-knock-out mouse by using the DNA construct of PLN gene designed by the result observed in the first step.
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Research Products
(12 results)
