| Project/Area Number |
07044289
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| Research Category |
Grant-in-Aid for international Scientific Research
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| Allocation Type | Single-year Grants |
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| Section | Joint Research |
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| Research Field |
内科学一般
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| Research Institution | Jichi Medical School |
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Principal Investigator |
KAWAKAMI Kiyoshi Jichi Medical School, Dept.Of Medicine, Associate Professor, 医学部, 助教授 (10161283)
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| Co-Investigator(Kenkyū-buntansha) |
BENZ Jr. Edward Johns Hopkins University, School of Medicine, Director, 医学部, 学部長
INGBAR David Minnesota University, School of Medicine, Associatet Professor, 医学部, 助教授
KOBAYASHI Makoto Jichi Medical School, Dept.Of Medicine, Assistant Professor, 医学部, 講師 (50254941)
MUTO Shigeaki Jichi Medical School, Dept.Of Medicine, Assistant Professor, 医学部, 講師 (40190855)
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| Project Period (FY) |
1995 – 1997
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| Keywords | Na, K-ATPase genes / cis elements / transacting fantor / transcription / osmolarity / hyperoxya / cardiocyte / nuclear extract |
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| Research Abstract |
1.In renal system, we applied cultured distal tubule cells and vascular smooth muscle cells and analyzed the response of Na, K-ATPase genes expression by trearment with corticosteroid hormones, high osmolarity or serum stimulation. Both a and b subunit gene were induced several fold by nortehrn blotting analysis. The induction was also observed when we transfected the reporter luciferase gene fused with the promote region of the Na, K-ATPase a1 and b1 subunit gene, indicating that the regulation was transcriptinal level.
2.Na, K-ATPase alpha3 subunit gene is transcribed in a few weeks after birth in rat heart. Using rat cardiocyte as a model system, we identified cis elements and transacting factors responsible for the gene. They are one NF-Y site and two Sp1/Sp3 sites. All three elements act as positive regulatory element and showed synergic activation. Protein occupancy on these elements were observed by in vivo footprinting analysis in cardiocyte specific manner.
3.The regulatory element of the beta1 subunit gene in response to hyperoxya was identified at -84 to -44 region. The two Sp1/Sp3 site in the region was not the resonsible element. For the analysis of the induction mechanism of the as subuunit gene during birth period, we established in vitro transcription system of the a1 subuit gene from adult and embyonic lung nuclear extracts. We identified negative regulatory element of the alpha1 subuit gene from -375 to -201. The cis element necessary for the full transcriptin activity was -155 to +31 in embryo while +66 to +92 was also necessary in adult.
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