| Project/Area Number |
08457285
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Kidney internal medicine
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| Research Institution | Tokyo Medical and Dental University |
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Principal Investigator |
INASE Naohiko (1997-1998) Tokyo Medical and Dental University, Scholl of Medicine Lectv, 医学部, 助手 (60262185)
佐々木 成 (1996) 東京医科歯科大学, 医学部, 助教授 (60170677)
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| Co-Investigator(Kenkyū-buntansha) |
UCHIDA Shinichi Tokyo Medical and Dental University, School of Medicine, Lecture, 医学部, 助手 (50262184)
SASAKI Sei Tokyo Medical and Dental University, Scholl of Medicine, Assoctate Prottese, 医学部, 助教授 (60170677)
稲瀬 直彦 東京医科歯科大学, 医学部, 助手 (60262185)
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| Project Period (FY) |
1996 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥8,200,000 (Direct Cost: ¥8,200,000)
Fiscal Year 1998: ¥2,700,000 (Direct Cost: ¥2,700,000)
Fiscal Year 1997: ¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 1996: ¥3,100,000 (Direct Cost: ¥3,100,000)
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| Keywords | AQP-2 / AQP-3 / CLC-K1 / CLC-K2 / Promoter / CRE / GATA / purine-rich sequence / 腎臓 / 輸送体 / 転写調節 / 核内タンパク / 遺伝子制御 |
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| Research Abstract |
Expression of aquaporin-2 (AQP2) is exclusively limited to kidney collecting duct cells, and this strictly limited expression could be mediated by transcription of the gene. The 5-flanking region of the AQP2 gene was characterized by a TATA box, two GATA consensus sequences, an AP-1 site, an AP-2 site, three E-boxes, and a cycle AMP-responsive element. A reporter gene assay performed in the 1st day of primary culture of IMCD cells showed that the 5' region up to -2.9 kb worked as a promoter. Deletion experiments showed that at least two regions, from -434 to -364 and from -153 to -84, contain negatively acting cis-elements. To evaluate the functional role of GATA motifs in AQP2 gene, we sought to isolate a GATA factor(s) expressed in collecting ducts. Two cDNAs encoding GATA factors were isolated from rat kidney (GATA-2 and -3). GATA motifs in the 5-flanking region of the AQP2 gene were functional cis-elements and that GATA-3 in collecting ducts may be one of the important regulators o
… More
f AQP-2 expression in vivo.
The rat CIC-K1 chloride channel is a kidney-specific member of the 010 chloride channel family found exclusively in the if-in ascending limb of Henle's loop in the kidney. To gain insight into the mechanism(s) of kidney-specific expression of CIC-K1, a genomic done that contains the 5'-flanking region of the rat CIC-K1 gene was isolated. The sequence of the proximal 5-flanking region contained an AP-3 site, a GRE, several AP-2 sites, and several E-boxes, but it lacked a TATA box.
Deletion analysis revealed that this cell-specific promoter activity in IM cells was still present in the construct containing 51 b p of-the 5'-flanking region but was lost in the -29 construct, dearly demonstrating that the-22 bp from -51 to -30 have a major role in the cell-specific activity of the CIC-K1-promoter. These 22 bp consist of purine-rich sequence (GGGGAGGGG-GAGGGGAG), and gel-retardation analysis demonstrated the existence of a specific protein(s) binding to this element in IM cells. These results suggest that the novel purine-rich element may play a key role in the activity of the CIC-K1 gene promoter. Less
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