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J. Biol. Chem., Vol. 279, Issue 40, 41830-41838, October 1, 2004

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Cloning and Characterization of Cytokeratins 8 and 19 in Adult Rat Striated Muscle

INTERACTION WITH THE DYSTROPHIN GLYCOPROTEIN COMPLEX^*

Jeanine A. Ursitti, Pervis C. Lee¶, Wendy G. Resneck¶, Minda M. McNally¶, Amber L. Bowman¶, Andrea O'Neill¶, Michele R. Stone¶, and Robert J. Bloch¶

From the University of Maryland Biotechnology Institute and the ^¶Department of Physiology, University of Maryland School of Medicine, Baltimore, Maryland 21201

We used degenerate primers for the amino- and carboxyl-terminal ends of the rod domains of intermediate filament proteins in reverse transcriptase-PCR experiments to identify and clone cytokeratins 8 and 19 (K8 and K19) from cardiac muscle of the adult rat. Northern blots showed that K8 has a 2.2-kb transcript and K19 has a 1.9-kb transcript in both adult cardiac and skeletal muscles. Immunolocalization of the cytokeratins in adult cardiac muscle with isoform-specific antibodies for K8 and K19 showed labeling at Z-lines within the muscle fibers and at Z-line and M-line domains at costameres at the sarcolemmal membrane. Dystrophin and K19 could be co-immunoprecipitated and co-purified from extracts of cardiac muscle, suggesting a link between the cytokeratins and the dystrophin-based cytoskeleton at the sarcolemma. Furthermore, transfection experiments indicate that K8 and K19 may associate with dystrophin through a specific interaction with its actin-binding domain. Consistent with this observation, the cytokeratins are disrupted at the sarcolemmal membrane of skeletal muscle of the mdx mouse that lacks dystrophin. Together these results indicate that at least two cytokeratins are expressed in adult striated muscle, where they may contribute to the organization of both the myoplasm and sarcolemma.

Received for publication, January 7, 2004 , and in revised form, June 24, 2004.

^* This work was supported by grants from the American Heart Association Mid-Atlantic Affiliate (to J. A. U.) and the Muscular Dystrophy Association (to R. J. B.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom all correspondence should be sent. Tel.: 410-706-4410; Fax: 410-706-8341; E-mail: jursitti{at}umaryland.edu.

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