title: Novel RNA-binding activity of MYF5 enhances Ccnd1/Cyclin D1 mRNA translation during myogenesis.
abstract: Skeletal muscle contains long multinucleated and contractile structures known as muscle fibers, which arise from the fusion of myoblasts into multinucleated myotubes during myogenesis. The myogenic regulatory factor (MRF) MYF5 is the earliest to be expressed during myogenesis and functions as a transcription factor in muscle progenitor cells (satellite cells) and myocytes. In mouse C2C12 myocytes, MYF5 is implicated in the initial steps of myoblast differentiation into myotubes. Here, using ribonucleoprotein immunoprecipitation (RIP) analysis, we discovered a novel function for MYF5 as an RNA-binding protein which associated with a subset of myoblast mRNAs. One prominent MYF5 target was Ccnd1 mRNA, which encodes the key cell cycle regulator CCND1 (Cyclin D1). Biotin-RNA pulldown, UV-crosslinking and gel shift experiments indicated that MYF5 was capable of binding the 3' untranslated region (UTR) and the coding region (CR) of Ccnd1 mRNA. Silencing MYF5 expression in proliferating myoblasts revealed that MYF5 promoted CCND1 translation and modestly increased transcription of Ccnd1 mRNA. Accordingly, overexpressing MYF5 in C2C12 cells upregulated CCND1 expression while silencing MYF5 reduced myoblast proliferation as well as differentiation of myoblasts into myotubes. Moreover, MYF5 silencing reduced myogenesis, while ectopically restoring CCND1 abundance partially rescued the decrease in myogenesis seen after MYF5 silencing. We propose that MYF5 enhances early myogenesis in part by coordinately elevating Ccnd1 transcription and Ccnd1 mRNA translation.
authors.affiliation: Laboratory of Genetics, National Institute on Aging, NIH, Baltimore, MD21224, USA amaresh.panda@nih.gov.
authors.forename: Amaresh C
authors.initials: AC
authors.lastname: Panda
authors.affiliation: Laboratory of Genetics, National Institute on Aging, NIH, Baltimore, MD21224, USA.
authors.forename: Kotb
authors.initials: K
authors.lastname: Abdelmohsen
authors.affiliation: Laboratory of Genetics, National Institute on Aging, NIH, Baltimore, MD21224, USA.
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Martindale
authors.affiliation: Translational Gerontology Branch, National Institute on Aging, NIH, Baltimore, MD 21224, USA Faculty of Veterinary Medicine, University of Teramo, Teramo, Italy.
authors.forename: Clara
authors.initials: C
authors.lastname: Di Germanio
authors.affiliation: Laboratory of Genetics, National Institute on Aging, NIH, Baltimore, MD21224, USA.
authors.forename: Xiaoling
authors.initials: X
authors.lastname: Yang
authors.affiliation: Laboratory of Genetics, National Institute on Aging, NIH, Baltimore, MD21224, USA.
authors.forename: Ioannis
authors.initials: I
authors.lastname: Grammatikakis
authors.affiliation: Laboratory of Genetics, National Institute on Aging, NIH, Baltimore, MD21224, USA.
authors.forename: Ji Heon
authors.initials: JH
authors.lastname: Noh
authors.affiliation: Laboratory of Genetics, National Institute on Aging, NIH, Baltimore, MD21224, USA.
authors.forename: Yongqing
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Laboratory of Genetics, National Institute on Aging, NIH, Baltimore, MD21224, USA.
authors.forename: Elin
authors.initials: E
authors.lastname: Lehrmann
authors.affiliation: Laboratory of Genetics, National Institute on Aging, NIH, Baltimore, MD21224, USA.
authors.forename: Dawood B
authors.initials: DB
authors.lastname: Dudekula
authors.affiliation: Laboratory of Genetics, National Institute on Aging, NIH, Baltimore, MD21224, USA.
authors.forename: Supriyo
authors.initials: S
authors.lastname: De
authors.affiliation: Laboratory of Genetics, National Institute on Aging, NIH, Baltimore, MD21224, USA.
authors.forename: Kevin G
authors.initials: KG
authors.lastname: Becker
authors.affiliation: Department of Biochemistry and Molecular Biology and Marlene and Stewart Greenebaum Cancer Center, University of Maryland School of Medicine, Baltimore, MD 21201, USA.
authors.forename: Elizabeth J
authors.initials: EJ
authors.lastname: White
authors.affiliation: Department of Biochemistry and Molecular Biology and Marlene and Stewart Greenebaum Cancer Center, University of Maryland School of Medicine, Baltimore, MD 21201, USA.
authors.forename: Gerald M
authors.initials: GM
authors.lastname: Wilson
authors.affiliation: Translational Gerontology Branch, National Institute on Aging, NIH, Baltimore, MD 21224, USA.
authors.forename: Rafael
authors.initials: R
authors.lastname: de Cabo
authors.affiliation: Laboratory of Genetics, National Institute on Aging, NIH, Baltimore, MD21224, USA.
authors.forename: Myriam
authors.initials: M
authors.lastname: Gorospe
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D002460:Cell Line; D049109:Cell Proliferation; D019938:Cyclin D1 / Q000235:genetics* / Q000378:metabolism; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D051379:Mice; D046228:Microarray Analysis; D024510:Muscle Development / Q000235:genetics*; D032446:Myoblasts; D051787:Myogenic Regulatory Factor 5 / Q000235:genetics* / Q000378:metabolism; D011485:Protein Binding; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction
chemical_list: C530105:Ccnd1 protein, mouse; C497418:Myf5 protein, mouse; D051787:Myogenic Regulatory Factor 5; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D019938:Cyclin D1
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