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J. Biol. Chem., Vol. 278, Issue 34, 31657-31666, August 22, 2003

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Epiplakin Gene Analysis in Mouse Reveals a Single Exon Encoding a 725-kDa Protein with Expression Restricted to Epithelial Tissues^*

Daniel Spazierer   ¶, Peter Fuchs  , Verena Pröll , Lubomir Janda , Susanne Oehler , Irmgard Fischer , Rudolf Hauptmann || and Gerhard Wiche  **

From the Vienna Biocenter, Institute of Biochemistry and Molecular Cell Biology, University of Vienna, A-1030 Vienna, Austria and the ^||Department of Exploratory Research, Boehringer Ingelheim Austria, A-1121 Vienna, Austria

Based on cDNA cloning and sequencing, human epiplakin has been classified as a member of the plakin protein family of cytolinkers. We report here the characterization of the mouse epiplakin gene locus and the isolation of full-length mouse epiplakin cDNA using BAC vectors. We found that the protein is encoded by a single remarkably large exon (>20 kb) that consists of a series of 0.8–1.5-kb-long DNA repeats, eight of which are virtually identical. Consequently, mouse epiplakin contains 16 plakin repeat domains, three more than reported for the human protein and eight more than predicted for the mouse protein based on the contig characterized by the Mouse Genome Sequencing Consortium. Using antibodies raised to a highly conserved repeating epiplakin sequence domain, we show that the protein in cells is expressed in its full length (725 kDa), and we provide evidence that the size of human epiplakin previously may have been underestimated. In addition we show on transcript and protein levels that epiplakin is restricted to epithelial tissues and that its gene maps to mouse chromosome 15 (human chromosome 8). This study lays the groundwork for future genetic approaches aimed at defining the biological role of this unique protein.

Received for publication, March 25, 2003 , and in revised form, June 5, 2003.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank^TM/EBI Data Bank with accession number(s) AY312170.

^* This work was supported by Grant SFB6-11 from the Austrian Science Research Fund. 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.

These authors contributed equally to this work.

^¶ Fellow of the International Ph.D. Program at the Vienna Biocenter, supported by Grant W1 from the Austrian Science Research Fund.

^** To whom correspondence should be addressed. E-mail: wiche{at}abc.univie.ac.at.

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