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Molecular and Cellular Biology, September 2007, p. 6163-6176, Vol. 27, No. 17
0270-7306/07/$08.00+0     doi:10.1128/MCB.01735-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

ROCK2 and Its Alternatively Spliced Isoform ROCK2m Positively Control the Maturation of the Myogenic Program{triangledown}

Michele Pelosi,1* Francesco Marampon,2 Bianca M. Zani,1,2 Sabrina Prudente,3 Emerald Perlas,1 Viviana Caputo,3 Luciano Cianetti,4 Valeria Berno,1 Shuh Narumiya,5 Shin W. Kang,6 Antonio Musarò,7 and Nadia Rosenthal1*

EMBL Mouse Biology Unit, Monterotondo (Roma), Italy,1 Department of Experimental Medicine, University of L'Aquila, L’Aquila, Italy,2 CSS-Mendel Institute, Roma, Italy,3 Department of Hematology, Oncology and Molecular Medicine, Istituto Superiore di Sanità, Roma, Italy,4 Department of Pharmacology, Faculty of Medicine, Kyoto University, Sakyo-ku, Kyoto, Japan,5 IRCCS Fondazione Santa Lucia, Roma, Italy,6 Department of Histology and Medical Embryology, University La Sapienza, Roma, Italy7

Received 14 September 2006/ Returned for modification 20 October 2006/ Accepted 10 June 2007

Signal transduction cascades involving Rho-associated kinases (ROCK), the serine/threonine kinases downstream effectors of Rho, have been implicated in the regulation of diverse cellular functions including cytoskeletal organization, cell size control, modulation of gene expression, differentiation, and transformation. Here we show that ROCK2, the predominant ROCK isoform in skeletal muscle, is progressively up-regulated during mouse myoblast differentiation and is highly expressed in the dermomyotome and muscle precursor cells of mouse embryos. We identify a novel and evolutionarily conserved ROCK2 splicing variant, ROCK2m, that is preferentially expressed in skeletal muscle and strongly up-regulated during in vivo and in vitro differentiation processes. The specific knockdown of ROCK2 or ROCK2m expression in C2C12 myogenic cells caused a significant and selective impairment of the expression of desmin and of the myogenic regulatory factors Mrf4 and MyoD. We demonstrate that in myogenic cells, ROCK2 and ROCK2m are positive regulators of the p42 and p44 mitogen-activated protein kinase-p90 ribosomal S6 kinase-eucaryotic elongation factor 2 intracellular signaling pathways and, thereby, positively regulate the hypertrophic effect elicited by insulin-like growth factor 1 and insulin, linking the multifactorial functions of ROCK to an important control of the myogenic maturation.

* Corresponding author. Mailing address: EMBL Mouse Biology Unit, Campus Buzzati-Traverso, via Ramarini 32, 00016 Monterotondo-Scalo, Roma, Italy. Phone: 39-06-90091241. Fax: 39-06-90091272. E-mail for Michele Pelosi: mpelosi{at}embl-monterotondo.it. E-mail for Nadia Rosenthal: rosenthal{at}embl-monterotondo.it

{triangledown} Published ahead of print on 2 July 2007.

Molecular and Cellular Biology, September 2007, p. 6163-6176, Vol. 27, No. 17
0270-7306/07/$08.00+0     doi:10.1128/MCB.01735-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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