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First published online 28 September 2004
doi: 10.1242/jcs.01406

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Xin repeats define a novel actin-binding motif

Dirk Pacholsky^1,*, Padmanabhan Vakeel^1,*, Mirko Himmel¹, Thomas Löwe¹, Theresia Stradal², Klemens Rottner², Dieter O. Fürst¹ and Peter F. M. van der Ven^1,

¹ Department of Cell Biology, Institute for Biochemistry and Biology, University of Potsdam, Karl-Liebknecht-Str. 24-25, 14476 Potsdam-Golm, Germany
² Department of Cell Biology, German Research Centre for Biotechnology, Mascheroder Weg 1, 38124 Braunschweig, Germany

Author for correspondence (e-mail: pvdven{at}rz.uni-potsdam.de)

Accepted 8 July 2004

Xin is a protein that is expressed during early developmental stages of cardiac and skeletal muscles. Immunolocalization studies indicated a peripheral localization in embryonic mouse heart, where Xin localizes with ß-catenin and N-cadherin. In adult tissues, Xin is found primarily in the intercalated discs of cardiomyocytes and the myotendinous junctions of skeletal muscle cells, both specialized attachment sites of the myofibrillar ends to the sarcolemma. A large part of the Xin protein consists of unique 16 amino acid repeats with unknown function. We have investigated the characteristics of the Xin repeats by transfection experiments and actin-binding assays and ascertained that, upon expression in cultured cells, these repeats bind to and stabilize the actin-based cytoskeleton. In vitro co-sedimentation assays with skeletal muscle actin indicated that they not only directly bind actin filaments, but also have the capability of arranging microfilaments into networks that sediment upon low-speed centrifugation. Very similar repeats were also found in `Xin-repeat protein 2' (XIRP2), a novel protein that seems to be expressed mainly in striated muscles. Human XIRP2 contains 28 Xin repeats with properties identical to those of Xin. We conclude that the Xin repeats define a novel, repetitive actin-binding motif present in at least two different muscle proteins. These Xin-repeat proteins therefore constitute the first two members of a novel family of actin-binding proteins.

Key words: Actin-binding proteins, Striated muscle, Repeats, Focal adhesions, Xin

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