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Am J Physiol Renal Physiol 279: F1132-F1138, 2000;
0363-6127/00 $5.00
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Vol. 279, Issue 6, F1132-F1138, December 2000

Translational regulation of Na-K-ATPase subunit mRNAs by glucocorticoids

Prasad Devarajan1 and Edward J. Benz Jr.2

1 Pediatric Nephrology, Yale University School of Medicine, New Haven 06520, and Albert Einstein College of Medicine, Bronx, New York 10467; and 2 Department of Medicine, Yale University School of Medicine, New Haven, Connecticut 06520; and Johns Hopkins School of Medicine, Baltimore, Maryland 21205

Glucocorticoids (GC) regulate Na-K-ATPase-subunit mRNA transcription. However, GC-induced increases in Na-K-ATPase activity are not always paralleled by changes in subunit mRNA abundance. We therefore examined posttranscriptional mechanisms of subunit gene regulation by GC. cDNA-derived mRNAs encoding alpha 1-, alpha 3-, and beta 1-subunits were tested for stability and translation efficiency in a cell-free lysate, in the presence of hydrocortisone (HC) or dexamethasone (Dex). No effect of HC on subunit mRNA stability was noted. Translation efficiency of alpha 1- and alpha 3-mRNAs, but not of beta 1-mRNA, was significantly increased by HC and Dex. Deletion of the 5'untranslated region (5'UT) of alpha 1-mRNA abolished this effect. Translation of a chimeric beta 1-mRNA, constructed by transposing the 5'UT of alpha 1 onto the coding region of beta 1, was enhanced by HC. Transposition of a putative steroid-modulatory element conserved in the 5'UT of all alpha  isoforms (ACAGGACCC) onto the coding region of beta 1-mRNA rendered it responsive to HC. A synthetic primer containing the ACAGGACCC sequence abolished the effect of HC on alpha 1- and chimeric beta 1-mRNAs. Our results indicate that GC can directly enhance Na-K-ATPase translation in vitro in a subunit-specific manner, via a putative GC-modulatory element situated in a predicted loop structure within the 5'UT of alpha -mRNAs.

mRNA stability; 5'untranslated region; hormonal regulation of sodium-potassium-adenosinetriphosphatase; glucocorticoid modulatory element; mRNA secondary structure.


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