Ubiquitin-proteasome-mediated degradation and synthesis of MyoD is modulated by alphaB-crystallin, a small heat shock protein, during muscle differentiation

Biochim Biophys Acta. 2010 Feb;1803(2):288-99. doi: 10.1016/j.bbamcr.2009.11.009. Epub 2009 Dec 11.

Abstract

alphaB-crystallin, a small heat shock protein, plays an important role in muscle homeostasis. It gets up-regulated during muscle differentiation and mice lacking alphaB-crystallin die prematurely with extensive muscle wastage. We have examined the role of alphaB-crystallin in muscle development using C2C12 myoblasts as a model system. Over-expression of alphaB-crystallin delays the muscle differentiation program significantly. C2C12 myoblasts over-expressing alphaB-crystallin (CRYAB-C2C12) display defect in cell-cycle exit upon induction of differentiation. During differentiation, CRYAB-C2C12 cells exhibit sustained level of cyclin D1 and delay in p21 and myogenin expression as compared to C2C12 cells. We find less accumulation of MyoD in CRYAB-C2C12 cells than in C2C12 cells. In vivo protein stability studies reveal faster ubiquitin-proteasome-mediated MyoD degradation in CRYAB-C2C12 cells (t(1/2)=1.42 h) than in C2C12 cells (t(1/2)=2.37 h). Immuno-precipitation experiments showed that MyoD gets ubiquitinated at earlier time points in CRYAB-C2C12 cells than in C2C12 cells. Our data reveal alterations in the synthesis and degradation of MyoD in CRYAB-C2C12 cells. The level of alphaB-crystallin as well as its Ser-59 phosphorylated form increases with increasing time of differentiation. Our studies show, inter alia, that alphaB-crystallin modulates myogenesis by altering MyoD level and provide an interesting insight in its role in myogenesis.

MeSH terms

  • Animals
  • Caspase 3 / metabolism
  • Cell Differentiation / physiology*
  • Cell Line
  • Cell Proliferation
  • Enzyme Activation
  • Heat-Shock Proteins, Small / metabolism
  • Mice
  • Muscle Development / physiology
  • Muscle, Skeletal / cytology
  • Muscle, Skeletal / physiology*
  • MyoD Protein / genetics
  • MyoD Protein / metabolism*
  • Myoblasts / cytology
  • Myoblasts / physiology
  • Proteasome Endopeptidase Complex / metabolism*
  • Ubiquitin / metabolism*
  • alpha-Crystallin B Chain / genetics
  • alpha-Crystallin B Chain / metabolism*

Substances

  • Heat-Shock Proteins, Small
  • MyoD Protein
  • Ubiquitin
  • alpha-Crystallin B Chain
  • Caspase 3
  • Proteasome Endopeptidase Complex