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J. Biol. Chem., Vol. 281, Issue 25, 16962-16970, June 23, 2006

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Carboxyl Terminus of Plakophilin-1 Recruits It to Plasma Membrane, whereas Amino Terminus Recruits Desmoplakin and Promotes Desmosome Assembly^*

From the Department of Oral Biology, University of Nebraska Medical Center College of Dentistry and Nebraska Center for Cellular Signaling, Omaha, Nebraska 68138

Plakophilins are armadillo repeat-containing proteins, initially identified as desmosomal plaque proteins that have subsequently been shown to also localize to the nucleus. Loss of plakophilin-1 is the underlying cause of ectodermal dysplasia/skin fragility syndrome, and skin from these patients exhibits desmosomes that are reduced in size and number. Thus, it has been suggested that plakophilin-1 plays an important role in desmosome stability and/or assembly. In this study, we used a cell culture system (A431DE cells) that expresses all of the proteins necessary to assemble a desmosome, except plakophilin-1. Using this cell line, we sought to determine the role of plakophilin-1 in de novo desmosome assembly. When exogenous plakophilin-1 was expressed in these cells, desmosomes were assembled, as assessed by electron microscopy and immunofluorescence localization of desmoplakin, into punctate structures. Deletion mutagenesis experiments revealed that amino acids 686–726 in the carboxyl terminus of plakophilin-1 are required for its localization to the plasma membrane. In addition, we showed that amino acids 1–34 in the amino terminus were necessary for subsequent recruitment of desmoplakin to the membrane and desmosome assembly.

Received for publication, January 19, 2006 , and in revised form, April 3, 2006.

^* This work was supported by National Institutes of Health Grant P20 RR018759 from the National Center for Research Resources (to J. K. W.). 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.

¹ To whom correspondence should be addressed: Dept. of Oral Biology University of Nebraska Medical Ctr. College of Dentistry, 987696 Nebraska Medical Ctr., Omaha, NE 68198-7696. Tel.: 402-559-3851; Fax: 402-559-3888; E-mail: jwahl{at}unmc.edu.

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