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• W4313371601 abstract "Despite its key role in potassium homeostasis, transcriptional control of the H + -K + -ATPase α 2 -subunit (HKα 2 ) gene in the collecting duct remains poorly characterized. cAMP increases H + -K + -ATPase activity in the collecting duct, but its role in activating HKα 2 transcription has not been explored. Previously, we demonstrated that the proximal 177 bp of the HKα 2 promoter confers basal collecting duct-selective expression. This region contains several potential cAMP/Ca 2+ -responsive elements (CRE). Accordingly, we examined the participation of CRE-binding protein (CREB) in HKα 2 transcriptional control in murine inner medullary collecting duct (mIMCD)-3 cells. Forskolin and vasopressin induced HKα 2 mRNA levels, and CREB overexpression stimulated the activity of HKα 2 promoter-luciferase constructs. Serial deletion analysis revealed that CREB inducibility was retained in a construct containing the proximal 100 bp of the HKα 2 promoter. In contrast, expression of a dominant negative inhibitor (A-CREB) resulted in 60% lower HKα 2 promoter-luciferase activity, suggesting that constitutive CREB participates in basal HKα 2 transcriptional activity. A constitutively active CREB mutant (CREB-VP16) strongly induced HKα 2 promoter-luciferase activity, whereas overexpression of CREBdLZ-VP16, which lacks the CREB DNA-binding domain, abolished this activation. In vitro DNase I footprinting and gel shift/supershift analysis of the proximal promoter with recombinant glutathione S-transferase (GST)-CREB-1 and mIMCD-3 cell nuclear extracts revealed sequence-specific DNA-CREB-1 complexes at −86/−60. Mutation at three CRE-like sequences within this region abolished CREB-1 DNA-binding activity and abrogated CREB-VP16 trans-activation of the HKα 2 promoter. In contrast, mutation of the neighboring −104/−94 κβ element did not alter CREB-VP16 trans-activation of the HKα 2 promoter. Thus CREB-1, binding to one or more CRE-like elements in the −86/−60 region, trans-activates the HKα 2 gene and may represent an important link between rapid and delayed effects of cAMP on HKα 2 activity." @default.
• W4313371601 created "2023-01-06" @default.
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• W4313371601 date "2004-10-01" @default.
• W4313371601 modified "2023-09-25" @default.
• W4313371601 title "CREB<i>trans</i>-activates the murine H⁺-K⁺-ATPase α₂-subunit gene" @default.
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• W4313371601 doi "https://doi.org/10.1152/ajpcell.00065.2004" @default.
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