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J. Biol. Chem., Vol. 281, Issue 22, 15352-15360, June 2, 2006

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Interaction of N-WASP with hnRNPK and Its Role in Filopodia Formation and Cell Spreading^*

Youngdong Yoo, Xiaoyang Wu, Coumaran Egile, Rong Li, and Jun-Lin Guan¹

From the Department of Molecular Medicine, Cornell University, Ithaca, New York 14853 and the Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115

N-WASP is a member of the WASP family of proteins, which play essential roles in actin dynamics during cell adhesion and migration. hnRNPK is a member of the heterogeneous nuclear ribonucleoprotein complex, which has also been implicated in the regulation of cell spreading. Here, we identify a direct interaction between N-WASP and hnRNPK. We show that this interaction is mediated by the N-terminal WH1 domain of N-WASP and the segment of hnRNPK containing its K interaction (KI) domain. Furthermore, these two proteins are co-localized at the cell periphery in the spreading initiation center during the early stage of cell spreading. We found that co-expression of hnRNPK with N-WASP reverses the stimulation of cell spreading by N-WASP, and this effect is correlated with hnRNPK binding to N-WASP. Expression of hnRNPK does not affect subcellular localization of N-WASP protein. However, co-expression of hnRNPK with N-WASP reduced filopodia formation stimulated by N-WASP in spreading cells. Together, these results identify hnRNPK as a new negative regulator of N-WASP and suggest that hnRNPK may regulate the initial stage of cell spreading by direct association with N-WASP in the spreading initiation center.

Received for publication, November 2, 2005 , and in revised form, March 28, 2006.

^* This work was supported by National Institutes of Health Grants GM48050 and HL73394 (to J.-L. G.). 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. Tel.: 607-253-3586; Fax: 607-253-3708; E-mail: jg19{at}cornell.edu.

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