title: Expression and subcellular localization of myogenic regulatory factors during the differentiation of skeletal muscle C2C12 myoblasts.
abstract: It is known that the MyoD family members (MyoD, Myf5, myogenin, and MRF4) play a pivotal role in the complex mechanism of skeletal muscle cell differentiation. However, fragmentary information on transcription factor-specific regulation is available and data on their post-transcriptional and post-translational behavior are still missing. In this work, we combined mRNA and protein expression analysis with their subcellular localization. Each myogenic regulator factor (MRF) revealed a specific mRNA trend and a protein quantitative analysis not overlapping, suggesting the presence of post-transcriptional mechanisms. In addition, each MRF showed a specific behavior in situ, characterized by a differentiation stage-dependent localization suggestive of a post-translational regulation also. Consistently with their transcriptional activity, immunogold electron microscopy data revealed MRFs distribution in interchromatin domains. Our results showed a MyoD and Myf5 contrasting expression profile in proliferating myoblasts, as well as myogenin and MRF4 opposite distribution in the terminally differentiated myotubes. Interestingly, MRFs expression and subcellular localization analysis during C2C12 cell differentiation stages showed two main MRFs regulation mechanisms: (i) the protein half-life regulation to modulate the differentiation stage-dependent transcriptional activity and (ii) the cytoplasmic retention, as a translocation process, to inhibit the transcriptional activity. Therefore, our results exhibit that MRFs nucleo-cytoplasmic trafficking is involved in muscle differentiation and suggest that, besides the MRFs expression level, also MRFs subcellular localization, related to their functional activity, plays a key role as a regulatory step in transcriptional control mechanisms.
authors.affiliation: Dipartimento di Scienze dell'Uomo, dell'Ambiente e della Natura, University of Urbino Carlo Bo, I-61029 Urbino, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Ferri
authors.forename: Elena
authors.initials: E
authors.lastname: Barbieri
authors.forename: Sabrina
authors.initials: S
authors.lastname: Burattini
authors.forename: Michele
authors.initials: M
authors.lastname: Guescini
authors.forename: Alessandra
authors.initials: A
authors.lastname: D'Emilio
authors.forename: Laura
authors.initials: L
authors.lastname: Biagiotti
authors.forename: Paolo
authors.initials: P
authors.lastname: Del Grande
authors.forename: Antonio
authors.initials: A
authors.lastname: De Luca
authors.forename: Vilberto
authors.initials: V
authors.lastname: Stocchi
authors.forename: Elisabetta
authors.initials: E
authors.lastname: Falcieri
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D005455:Fluorescent Antibody Technique; D051379:Mice; D018482:Muscle, Skeletal / Q000166:cytology* / Q000378:metabolism; D032446:Myoblasts / Q000378:metabolism*; D018006:Myogenic Regulatory Factors / Q000032:analysis* / Q000378:metabolism*
chemical_list: D018006:Myogenic Regulatory Factors