Insulin regulates protein kinase CbetaII expression through enhanced exon inclusion in L6 skeletal muscle cells. A novel mechanism of insulin- and insulin-like growth factor-i-induced 5' splice site selection.
The protein kinase Cbeta (PKCbeta) gene encodes two isoforms, PKCbetaI and PKCbetaII, as a result of alternative splicing. The unique mechanism that underlies insulin-induced alternative splicing of PKCbeta pre-mRNA was examined in L6 myotubes. Mature PKCbetaII mRNA and protein rapidly increased >3-fold following acute insulin treatment, while PKCbetaI mRNA and protein levels remained unchanged. Mature PKCbetaII mRNA resulted from inclusion of the PKCbetaII-specific exon rather than from selection of an alternative polyadenylation site. Increased PKCbetaII expression was also not likely accounted for by transcriptional activation of the gene or increased stabilization of the PKCbetaII mRNA, and suggest that PKCbetaII expression is regulated primarily at the level of alternative splicing. Insulin effects on exon inclusion were observed as early as 15 min after insulin treatment; by 20 min, a new 5'-splice site variant of PKCbetaII was also observed. After 30 min, the longer 5'-splice site variant became the predominate species through activation of a downstream 5' splice site. Similar results were obtained using IGF-I. Although the role of this new PKCbetaII mRNA species is presently unknown, inclusion of either PKCbetaII-specific exon results in the same PKCbetaII protein.
Chalfant
C E
CE
Department of Biochemistry and Molecular Biology, University of South Florida College of Medicine, Tampa, Florida 33612, USA.
Watson
J E
JE
Bisnauth
L D
LD
Kang
J B
JB
Patel
N
N
Obeid
L M
LM
Eichler
D C
DC
Cooper
D R
DR
D017398:Alternative Splicing; D000818:Animals; D002460:Cell Line; D005091:Exons; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects*; D007328:Insulin / Q000494:pharmacology*; D007334:Insulin-Like Growth Factor I / Q000494:pharmacology*; D007527:Isoenzymes / Q000235:genetics*; D018482:Muscle, Skeletal / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology; D011493:Protein Kinase C / Q000235:genetics*; D064546:Protein Kinase C beta; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats
D007328:Insulin; D007527:Isoenzymes; D012333:RNA, Messenger; D007334:Insulin-Like Growth Factor I; D011493:Protein Kinase C; D064546:Protein Kinase C beta