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• W2001520624 abstract "Abstract On the basis of paradigms in development wherein discrete transcriptional events are pivotal regulatory steps, we tested the hypothesis that transcriptional sodium (Na + )–response mechanisms are involved in in vivo Na + -induced responses relevant to normal (homeostatic) and pathophysiological (salt-sensitive hypertension) conditions. We used Na,K-ATPase α-subunit genes as molecular probes and the Na + ionophore monensin to induce a dose-specific incremental increase in [Na + ] i in rat A10 embryonic aortic smooth muscle cells. RNA blot analysis of rat A10 cells revealed a dose-specific (0.022 to 30 μmol/L monensin) upregulation of α 1 -, α 2 -, and β 1 -subunit Na,K-ATPase RNA levels. Control β-actin and α-tropomyosin RNA levels did not change. With the use of chloramphenicol acetyltransferase (CAT) as reporter gene, CAT assays of rat α 1 [−1288]CAT and human α 2 [−798]CAT promoter constructs exhibited induction of CAT activity in monensin (10 μmol/L)–treated A10 cells compared with untreated A10 cells. Promoter deletion constructs for rat α 1 [−1288]CAT defined a positive Na + -response regulatory region within −358 to −169 that is distinct from the basal transcriptional activation region of −155 to −49 previously defined. Similarly, a positive Na + -response regulatory region is delimited to within −301 in the human α 2 Na,K-ATPase 5′ flanking region. Analysis of transgenic TgHα 2 [−798]CAT rats demonstrated sodium activation of human α 2 [−798]CAT transgene expression in aorta parallel to observations made in rat A10 aortic tissue culture cells. Southwestern blot analysis of nuclear extracts from monensin (10 μmol/L)–treated and control untreated A10 cells revealed a nuclear DNA binding protein (approximately 95 kD) that is upregulated by increased [Na + ] i . These data provide initial characterization of a transcriptional Na + -response mechanism delimiting a positive Na + -response regulatory region in two target genes (α 1 and α 2 Na,K-ATPase) as well as detection of a Na + -response nuclear DNA binding protein. The in vitro data are corroborated by in vivo experimental and transgenic promoter expression studies, thus validating the biological relevance of the observations." @default.
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• W2001520624 date "1997-08-01" @default.
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• W2001520624 title "Characterization of a Sodium-Response Transcriptional Mechanism" @default.
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• W2001520624 doi "https://doi.org/10.1161/01.hyp.30.2.191" @default.
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