Post-transcriptional regulation of Na+/glucose cotransporter (SGTL1) gene expression in LLC-PK1 cells. Increased message stability after cyclic AMP elevation or differentiation inducer treatment.
Abstract
We have further investigated the molecular basis of increased differentiation-regulated expression of SGTL1, a Na+/glucose cotransporter, in the renal epithelial cell line LLC-PK1. Treatment of confluent monolayers either with the differentiation inducer hexamethylene bisacetamide (HMBA) or with cyclic AMP-elevating agents promoted increased levels of the SGLT1 mRNA, the immunodetectable 75-kDa cotransporter subunit, and the transport activity. Two molecular species of SGLT1 mRNA (2.2 and 3.9 kilobases (kb)) are transcribed from the same gene in LLC-PK1 cells and differ only in the length of the 3'-untranslated region. The larger transcript is less stable (t1/2 = 2 h) than the smaller one (t1/2 = 10 h) in control, confluent monolayers. The 3.9-kb species was stabilized from degradation after either cyclic AMP elevation (t1/2 = 14 h) or HMBA addition (t1/2 = 8 h), with negligible effects on the stability of the 2.2-kb species (t1/2 = 11 h). Inhibition of translation by cycloheximide resulted in a 10-fold increase in the t1/2 of the 3.9-kb transcript and a 2-fold increase in that of the 2.2-kb species in control monolayers. Our results demonstrate that post-transcriptional regulation of message stability plays a major role in dif...Continue Reading
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