title: Inositide-dependent phospholipase C signaling mimics insulin in skeletal muscle differentiation by affecting specific regions of the cyclin D3 promoter.
abstract: Our main goal in this study was to investigate the role of phospholipase C (PLC) beta(1) and PLCgamma(1) in skeletal muscle differentiation and the existence of potential downstream targets of their signaling activity. To examine whether PLC signaling can modulate the expression of cyclin D3, a target of PLCbeta(1) in erythroleukemia cells, we transfected C2C12 cells with expression vectors containing PLCbeta(1) or PLCgamma(1) cDNA and with small interfering RNAs from regions of the PLCbeta(1) or PLCgamma(1) gene and followed myogenic differentiation in this well-established cell system. Intriguingly, overexpressed PLCbeta(1) and PLCgamma(1) were able to mimic insulin induction of both cyclin D3 and muscle differentiation. By knocking down PLCbeta(1) or PLCgamma(1) expression, C2C12 cells almost completely lost the increase in cyclin D3, and the differentiation program was down-regulated. To explore the induction of the cyclin D3 gene promoter during this process, we used a series of 5'-deletions of the 1.68-kb promoter linked to a reporter gene and noted a 5-fold augmentation of promoter activity upon insulin stimulation. These constructs were also cotransfected with PLCbeta(1) or PLCgamma(1) cDNAs and small interfering RNAs, respectively. Our data indicate that PLCbeta(1) or PLCgamma(1) signaling is capable of acting like insulin in regard to both the myogenic differentiation program and cyclin D3 up-regulation. Taken together, this is the first study that hints at cyclin D3 as a target of PLCbeta(1) and PLCgamma(1) during myogenic differentiation in vitro and implies that up-regulation of these enzymes is sufficient to mimic the actions of insulin in this process.
authors.affiliation: Cellular Signalling Laboratory, Department of Human Anatomical Sciences, University of Bologna, Via Irnerio 48, 40126 Bologna, Italy.
authors.forename: Irene
authors.initials: I
authors.lastname: Faenza
authors.forename: Giulia
authors.initials: G
authors.lastname: Ramazzotti
authors.forename: Alberto
authors.initials: A
authors.lastname: Bavelloni
authors.forename: Roberta
authors.initials: R
authors.lastname: Fiume
authors.forename: Gian Carlo
authors.initials: GC
authors.lastname: Gaboardi
authors.forename: Matilde Y
authors.initials: MY
authors.lastname: Follo
authors.forename: R Stewart
authors.initials: RS
authors.lastname: Gilmour
authors.forename: Alberto M
authors.initials: AM
authors.lastname: Martelli
authors.forename: Katya
authors.initials: K
authors.lastname: Ravid
authors.forename: Lucio
authors.initials: L
authors.lastname: Cocco
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D056743:Cyclin D3; D016213:Cyclins / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D007150:Immunohistochemistry; D007328:Insulin / Q000502:physiology*; D007527:Isoenzymes / Q000378:metabolism / Q000502:physiology*; D051379:Mice; D024510:Muscle Development / Q000187:drug effects; D018482:Muscle, Skeletal / Q000201:enzymology*; D018008:Myogenin / Q000502:physiology; D054799:Phospholipase C beta; D051966:Phospholipase C gamma / Q000378:metabolism / Q000502:physiology; D011401:Promoter Regions, Genetic; D034741:RNA, Small Interfering / Q000494:pharmacology; D015398:Signal Transduction; D010738:Type C Phospholipases / Q000378:metabolism / Q000502:physiology*
chemical_list: C534696:CCND3 protein, human; C534697:Ccnd3 protein, mouse; D056743:Cyclin D3; D016213:Cyclins; D007328:Insulin; D007527:Isoenzymes; C503801:Myog protein, mouse; D018008:Myogenin; D034741:RNA, Small Interfering; D010738:Type C Phospholipases; D054799:Phospholipase C beta; C518034:Plcb1 protein, mouse; D051966:Phospholipase C gamma