Interaction of the Nck Adapter Protein with p21-activated Kinase (PAK1)

doi:10.1074/jbc.271.42.25746

Interaction of the Nck Adapter Protein with p21-activated Kinase (PAK1)*

From the ^‡ Department of Immunology and Department of Cell Biology, The Scripps Research Institute, La Jolla, California 92037,
^§ Fox Chase Cancer Center, Philadelphia, Pennsylvania 19111, and
^¶ Department of Biochemistry and Molecular Biology and Walther Oncology Center, Indiana University, Indianapolis, Indiana 46202

Abstract

The p21-activated kinases (PAKs) link G protein-coupled receptors and growth factor receptors (S. Dharmawardhane, R. H. Daniels, and G. M. Bokoch, submitted for publication) to activation of MAP kinase cascades and to cytoskeletal reorganization (M. A. Sells, U. G. Knaus, D. Ambrose, S. Bagrodia, G. M. Bokoch, and J. Chernoff, submitted for publication). The proteins that interact with PAK to mediate its cellular effects and to couple it to upstream receptors are unknown. We describe here a specific interaction of the Nck adapter molecule with PAK1 both in vitro and in vivo. PAK1 and Nck associate in COS-7 and Swiss 3T3 cells constitutively, but this interaction is strengthened upon platelet-derived growth factor receptor stimulation. We show that Nck binds to PAK1 through its second Src homology 3 (SH3) domain, while PAK1 interacts with Nck via the first proline-rich SH3 binding motif at its amino terminus. The interaction of active PAK1 with Nck leads to the phosphorylation of Nck at multiple sites. Association of Nck with PAK1 may serve to link this important regulatory kinase to cell activation by growth factor receptors.

Footnotes

↵* This work was supported by United States Public Health Service Grants GM39434 (to G. M. B.), CA63139 (to L. A. Q.), and AI35947 (to U. G. K.). The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
↵¹ The abbreviations used are:

SH2

Src homology 2

SH3

Src homology 3

EGF

epidermal growth factor

PDGF

platelet-derived growth factor

PKA

cAMP-dependent protein kinase

PAK

p21-activated kinase

GST

glutathione S-transferase

GTPγS

guanosine 5′-3-O(thio)triphosphate

aa

amino acids.
↵² M. A. Sells, U. G. Knaus, D. Ambrose, S. Bagrodia, G. M. Bokoch, and J. Chernoff, submitted for publication.
↵³ S. Dharmawardhane, R. H. Daniels, and G. M. Bokoch, submitted for publication.
↵⁴ R. H. Daniels and G. M. Bokoch, unpublished observations.
- Received July 30, 1996.
- Revision received August 30, 1996.