Signal-dependent nuclear export of a histone deacetylase regulates muscle differentiation

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Abstract

Members of the myocyte enhancer factor-2 (MEF2) family of transcription factors associate with myogenic basic helix–loop–helix transcription factors such as MyoD to activate skeletal myogenesis1. MEF2 proteins also interact with the class II histone deacetylases HDAC4 and HDAC5, resulting in repression of MEF2-dependent genes2,3,4. Execution of the muscle differentiation program requires release of MEF2 from repression by HDACs, which are expressed constitutively in myoblasts and myotubes5. Here we show that HDAC5 shuttles from the nucleus to the cytoplasm when myoblasts are triggered to differentiate. Calcium/calmodulin-dependent protein kinase (CaMK) signalling, which stimulates myogenesis5 and prevents formation of MEF2–HDAC complexes4, also induces nuclear export of HDAC4 and HDAC5 by phosphorylation of these transcriptional repressors. An HDAC5 mutant lacking two CaMK phosphorylation sites is resistant to CaMK-mediated nuclear export and acts as a dominant inhibitor of skeletal myogenesis, whereas a cytoplasmic HDAC5 mutant is unable to block efficiently the muscle differentiation program. Our results highlight a mechanism for transcriptional regulation through signal- and differentiation-dependent nuclear export of a chromatin-remodelling enzyme, and suggest that nucleo-cytoplasmic trafficking of HDACs is involved in the control of cellular differentiation.

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Figure 1: Shuttling of HDAC5 from the nucleus to the cytoplasm during myogenic differentiation.
Figure 2: HDAC5 is excluded from the nucleus in cells expressing activated forms of CaMK.
Figure 3: CaMK signalling stimulates nuclear export of HDAC5.
Figure 4: Identification of CaMK target sites in HDAC5.
Figure 5: Regulation of myogenesis by HDAC5 nuclear export.

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Acknowledgements

We thank M. Cobb, J. Han, R. Lin, G.S. McKnight, A. Means, S. O'Keefe, S. Schreiber, T. Chatila and J. Woodgett for expression plasmids; R. Prives for anti-MEF2 antisera; and M. Yoshida for leptomycin B. We are grateful to A. Tizenor for graphics, and J. Page and W. Simpson for editorial assistance. E.N.O. was supported by grants from NIH, the Robert A. Welch Foundation, the D. W. Reynolds Foundation and Myogen, Inc. T.A.M. is a Pfizer fellow of The Life Sciences Research Foundation.

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Correspondence to Eric N. Olson.

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