Requirement of Nck adaptors for actin dynamics and cell migration stimulated by platelet-derived growth factor B

Raymond and Beverly Sackler Laboratory of Genetics and Molecular Medicine, Department of Genetics and Developmental Biology and Center for Cell Analysis and Modeling, University of Connecticut Health Center, Farmington, CT 06030, USA.
Proceedings of the National Academy of Sciences (Impact Factor: 9.67). 07/2006; 103(25):9536-41. DOI: 10.1073/pnas.0603786103
Source: PubMed


The Nck family of Src homology (SH) 2/SH3 domain adaptors functions to link tyrosine phosphorylation induced by extracellular signals with downstream regulators of actin dynamics. We investigated the role of mammalian Nck adaptors in signaling from the activated platelet-derived growth factor (PDGF) receptor (PDGFbetaR) to the actin cytoskeleton. We report here that Nck adaptors are required for cytoskeletal reorganization and chemotaxis stimulated by PDGF-B. Analysis of tyrosine-phosphorylated proteins demonstrated that Crk-associated substrate (p130(Cas)), not the activated PDGFbetaR itself, is the major Nck SH2 domain-binding protein in PDGF-B-stimulated cells. Both Nck- and p130(Cas)-deficient cells fail to display cytoskeletal rearrangements, including the formation of membrane ruffles and the disassembly of actin bundles, typically shown by their WT counterparts in response to PDGF-B. Furthermore, Nck and p130(Cas) colocalize in phosphotyrosine-enriched membrane ruffles induced by PDGF-B in NIH 3T3 cells. These results suggest that Nck adaptors play an essential role in linking the activated PDGFbetaR with actin dynamics through a pathway that involves p130(Cas).

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Available from: Nah-Young Shin, Nov 01, 2014
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    • "These results suggest that N-WASP and Arp2/3 complex are part of a multiprotein assembly important for the generation of PDGFAA-induced dorsal ruffles. A similar correlation between Nck and the formation of dorsal ruffles has been observed [52]. These observations clearly put Nck and its binding partners in a common pathway in the formation of dorsal ruffles. "
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    ABSTRACT: The spatial distribution of signals downstream from receptor tyrosine kinases (RTKs) or G-protein coupled receptors (GPCR) regulates fundamental cellular processes that control cell migration and growth. Both pathways rely significantly on actin cytoskeleton reorganization mediated by nucleation-promoting factors such as the WASP-(Wiskott-Aldrich Syndrome Protein) family. WIP (WASP Interacting Protein) is essential for the formation of a class of polarised actin microdomain, namely dorsal ruffles, downstream of the RTK for PDGF (platelet-derived growth factor) but the underlying mechanism is poorly understood. Using lentivirally-reconstituted WIP-deficient murine fibroblasts we define the requirement for WIP interaction with N-WASP (neural WASP) and Nck for efficient dorsal ruffle formation and of WIP-Nck binding for fibroblast chemotaxis towards PDGF-AA. The formation of both circular dorsal ruffles in PDGF-AA-stimulated primary fibroblasts and lamellipodia in CXCL13-treated B lymphocytes are also compromised by WIP-deficiency. We provide data to show that a WIP-Nck signalling complex interacts with RTK to promote polarised actin remodelling in fibroblasts and provide the first evidence for WIP involvement in the control of migratory persistence in both mesenchymal (fibroblast) and amoeboid (B lymphocytes) motility.
    Full-text · Article · Aug 2013 · PLoS ONE
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    • "In our analysis, we find a genome-wide significant association of NCK2 gene on chromosome 2 with opiates dependence in African-origin men at both the SNP and gene levels. NCK2 is a member of NCK family of adaptor proteins, which is associated with tyrosine-phosphorylated growth factor receptors of their cellular substrates [54]. However, to the best of our knowledge, NCK2 has not been reported to be associated with any drug addiction outcomes in humans. "
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    • "The role of CAS in cell migration is linked to its capacity to recruit a range of signaling molecules including the adaptor proteins Crk and Nck (Klemke et al., 1998; Rivera et al., 2006). To assess whether Crk and Nck were associated with the induction of EGFRmediated carcinoma cell migration, lysates from FG cells treated with or without EGF were subjected to immunoprecipitation with anti-CAS followed by immunoblotting for Nck1, Nck2, CrkL and CrkI. "
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