Control of insulin gene expression in pancreatic beta-cells and in an insulin-producing cell line, RIN-5F cells. II. Regulation of insulin mRNA stability.
The half-life of insulin mRNA at various glucose concentrations was determined by filter hybridization techniques in isolated rat islets incubated with 3H-labeled uridine followed by a chase incubation at 3.3 or 17 mM glucose. High glucose induced a greater stabilization of insulin mRNA than of other poly(A) + RNAs or total cellular RNA. In RIN-5F insulinoma cells, an insulin-producing cell line, cholera toxin, but not glucose, induced a stabilization of insulin mRNA. After 24 h of culture of islets with actinomycin D or alpha-amanitin at several glucose concentrations, insulin mRNA content was decreased in comparison to controls only at higher glucose concentrations. The biosynthesis of islet proteins other than insulin was strongly decreased by actinomycin D at all glucose concentrations. Insulin biosynthesis was inhibited proportionately to the observed decreases in insulin mRNA content. We conclude that inhibition of insulin mRNA degradation is an important component in increasing the insulin mRNA content in response to glucose, thereby augmenting the effects of glucose stimulation on insulin gene transcription (5). This stabilization may be partly mediated by cAMP as evidenced by the similar responses to cholera toxin in the RIN-5F cells. Furthermore, the results of experiments with actinomycin D suggest that the degradation of insulin mRNA may require the continuous production of a factor(s) which could be either RNA or protein in nature.
D000546:Amanitins / Q000494:pharmacology; D000818:Animals; D002460:Cell Line; D003609:Dactinomycin / Q000494:pharmacology; D005786:Gene Expression Regulation; D005947:Glucose / Q000494:pharmacology; D007328:Insulin / Q000096:biosynthesis / Q000235:genetics*; D007515:Islets of Langerhans / Q000378:metabolism*; D012333:RNA, Messenger / Q000032:analysis*; D051381:Rats
D000546:Amanitins; D007328:Insulin; D012333:RNA, Messenger; D003609:Dactinomycin; D005947:Glucose