Cyclin E-cdk2 phosphorylation promotes late G1-phase degradation of MyoD in muscle cells.
Proliferating myoblasts already express MyoD before the induction of differentiation. Overexpression of MyoD in normal and transformed cell lines was shown to block cells from entering S phase, suggesting that the MyoD growth suppressive effect must be tightly controlled in growing myoblasts. Here we show that during G1 phase, but not in G2, MyoD abundance is down-regulated by the ubiquitin-proteasome pathway through phosphorylation of serine 200. Roscovitine, a specific inhibitor of cyclin-Cdk2 complexes, prevents both phosphorylation and degradation of MyoD in G1. Inhibition of the ubiquitin-dependent proteasome pathway by MG132 results in stabilization of MyoD-wt, with little effect on a MyoD mutant where serine 200 is replaced by an alanine. Our results show that MyoD Ser200 is the substrate for phosphorylation by cyclin E-Cdk2 stimulating its degradation by the ubiquitin-proteasome system which controls MyoD levels in G1. Phosphorylation/degradation of MyoD at the end of G1 thus represents the regulatory checkpoint in growing myoblasts allowing progression into S phase in a manner similar to the recently examplified cdk2-phosphorylation/degradation of p27(Kip1).
Tintignac
L A
LA
Laboratoire de Génétique Oncologique, Institut Gustave Roussy, Villejuif, 94805, France.
Leibovitch
M P
MP
Kitzmann
M
M
Fernandez
A
A
Ducommun
B
B
Meijer
L
L
Leibovitch
S A
SA
D000818:Animals; D042846:CDC2-CDC28 Kinases; D019927:Cyclin E / Q000378:metabolism*; D051357:Cyclin-Dependent Kinase 2; D018844:Cyclin-Dependent Kinases / Q000378:metabolism*; D003546:Cysteine Endopeptidases / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005347:Fibroblasts / Q000166:cytology; D016193:G1 Phase / Q000187:drug effects / Q000502:physiology*; D051379:Mice; D009097:Multienzyme Complexes / Q000378:metabolism; D018485:Muscle Fibers, Skeletal / Q000166:cytology* / Q000201:enzymology; D018482:Muscle, Skeletal / Q000166:cytology; D017570:MyoD Protein / Q000378:metabolism*; D010766:Phosphorylation; D046988:Proteasome Endopeptidase Complex; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D011687:Purines / Q000494:pharmacology; D000077546:Roscovitine; D016196:S Phase / Q000187:drug effects / Q000502:physiology; D012694:Serine; D013379:Substrate Specificity; D014452:Ubiquitins / Q000378:metabolism
D019927:Cyclin E; D004791:Enzyme Inhibitors; D009097:Multienzyme Complexes; D017570:MyoD Protein; D011687:Purines; D014452:Ubiquitins; D000077546:Roscovitine; D012694:Serine; D017346:Protein Serine-Threonine Kinases; D042846:CDC2-CDC28 Kinases; C495893:Cdk2 protein, mouse; D051357:Cyclin-Dependent Kinase 2; D018844:Cyclin-Dependent Kinases; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex