title: Fam65b is important for formation of the HDAC6-dysferlin protein complex during myogenic cell differentiation.
abstract: Previously, we identified family with sequence similarity 65, member B (Fam65b), as a protein transiently up-regulated during differentiation and fusion of human myogenic cells. Silencing of Fam65b expression results in severe reduction of myogenin expression and consequent lack of myoblast fusion. The molecular function of Fam65b and whether misregulation of its expression could be causative of muscle diseases are unknown. Protein pulldowns were used to identify Fam65b-interacting proteins in differentiating human muscle cells and regenerating muscle tissue. In vitro, human muscle cells were treated with histone-deacetylase (HDAC) inhibitors, and expression of Fam65b and interacting proteins was studied. Nontreated cells were used as controls. In vivo, expression of Fam65b was down-regulated in developing zebrafish to determine the effects on muscle development. Fam65b binds to HDAC6 and dysferlin, the protein mutated in limb girdle muscular dystrophy 2B. The tricomplex Fam65b-HDAC6-dysferlin is transient, and Fam65b expression is necessary for the complex to form. Treatment of myogenic cells with pan-HDAC or HDAC6-specific inhibitors alters Fam65b expression, while dysferlin expression does not change. Inhibition of Fam65b expression in developing zebrafish results in abnormal muscle, with low birefringence, tears at the myosepta, and increased embryo lethality. Fam65b is an essential component of the HDAC6-dysferlin complex. Down-regulation of Fam65b in developing muscle causes changes consistent with muscle disease.-Balasubramanian, A., Kawahara, G., Gupta, V. A., Rozkalne, A., Beauvais, A., Kunkel, L. M., Gussoni, E. Fam65b is important for formation of the HDAC6-dysferlin protein complex during myogenic cell differentiation.
authors.affiliation: Program in Genomics, Division of Genetics and.
authors.forename: Anuradha
authors.initials: A
authors.lastname: Balasubramanian
authors.affiliation: Program in Genomics, Division of Genetics and.
authors.forename: Genri
authors.initials: G
authors.lastname: Kawahara
authors.affiliation: Program in Genomics, Division of Genetics and.
authors.forename: Vandana A
authors.initials: VA
authors.lastname: Gupta
authors.affiliation: Program in Genomics, Division of Genetics and.
authors.forename: Anete
authors.initials: A
authors.lastname: Rozkalne
authors.affiliation: Regenerative Medicine Program, Ottawa Hospital Research Institute, Ottawa, Ontario, Canada; and.
authors.forename: Ariane
authors.initials: A
authors.lastname: Beauvais
authors.affiliation: Program in Genomics, Division of Genetics and Manton Center for Orphan Disease Research, Boston Children's Hospital, Boston, Massachusetts, USA; Harvard Medical School, Boston, Massachusetts, USA.
authors.forename: Louis M
authors.initials: LM
authors.lastname: Kunkel
authors.affiliation: Program in Genomics, Division of Genetics and Harvard Medical School, Boston, Massachusetts, USA gussoni@enders.tch.harvard.edu.
authors.forename: Emanuela
authors.initials: E
authors.lastname: Gussoni
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D015815:Cell Adhesion Molecules; D002454:Cell Differentiation / Q000235:genetics*; D002478:Cells, Cultured; D015536:Down-Regulation / Q000235:genetics; D000073939:Dysferlin; D000073864:Histone Deacetylase 6; D006655:Histone Deacetylases / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D032342:Muscle Cells / Q000378:metabolism*; D024510:Muscle Development / Q000235:genetics*; D009124:Muscle Proteins / Q000235:genetics / Q000378:metabolism*; D018482:Muscle, Skeletal / Q000378:metabolism; D009135:Muscular Diseases / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding / Q000235:genetics; D011506:Proteins / Q000235:genetics* / Q000378:metabolism*; D016415:Sequence Alignment; D014404:Tubulin / Q000235:genetics / Q000378:metabolism; D015027:Zebrafish
chemical_list: D015815:Cell Adhesion Molecules; C114839:DYSF protein, human; D000073939:Dysferlin; D008565:Membrane Proteins; D009124:Muscle Proteins; D011506:Proteins; C104952:RIPOR2 protein, human; D014404:Tubulin; C433811:HDAC6 protein, human; D000073864:Histone Deacetylase 6; D006655:Histone Deacetylases
keywords: acetylation; muscular dystrophy; myoblast; zebrafish