Paxillin Kinase Linker (PKL) Regulates Vav2 Signaling During Cell Spreading and Migration

Overview

Journal Mol Biol Cell

Specialties Cell Biology
Molecular Biology

Date 2013 Apr 26

PMID 23615439

Citations 6

Authors

Matthew C Jones

Kazuya Machida

Bruce J Mayer

Christopher E Turner

Affiliations

Soon will be listed here.

Abstract

The Rho family of GTPases plays an important role in coordinating dynamic changes in the cell migration machinery after integrin engagement with the extracellular matrix. Rho GTPases are activated by guanine nucleotide exchange factors (GEFs) and negatively regulated by GTPase-activating proteins (GAPs). However, the mechanisms by which GEFs and GAPs are spatially and temporally regulated are poorly understood. Here the activity of the proto-oncogene Vav2, a GEF for Rac1, RhoA, and Cdc42, is shown to be regulated by a phosphorylation-dependent interaction with the ArfGAP PKL (GIT2). PKL is required for Vav2 activation downstream of integrin engagement and epidermal growth factor (EGF) stimulation. In turn, Vav2 regulates the subsequent redistribution of PKL and the Rac1 GEF β-PIX to focal adhesions after EGF stimulation, suggesting a feedforward signaling loop that coordinates PKL-dependent Vav2 activation and PKL localization. Of interest, Vav2 is required for the efficient localization of PKL and β-PIX to the leading edge of migrating cells, and knockdown of Vav2 results in a decrease in directional persistence and polarization in migrating cells, suggesting a coordination between PKL/Vav2 signaling and PKL/β-PIX signaling during cell migration.

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References

Webb D, Parsons J, Horwitz A . Adhesion assembly, disassembly and turnover in migrating cells -- over and over and over again. Nat Cell Biol. 2002; 4(4):E97-100. DOI: 10.1038/ncb0402-e97. View

Abe K, Rossman K, Liu B, Ritola K, Chiang D, Campbell S . Vav2 is an activator of Cdc42, Rac1, and RhoA. J Biol Chem. 2000; 275(14):10141-9. DOI: 10.1074/jbc.275.14.10141. View

Yu J, Deakin N, Turner C . Paxillin-kinase-linker tyrosine phosphorylation regulates directional cell migration. Mol Biol Cell. 2009; 20(22):4706-19. PMC: 2777101. DOI: 10.1091/mbc.e09-07-0548. View

West K, Zhang H, Brown M, Nikolopoulos S, Riedy M, Horwitz A . The LD4 motif of paxillin regulates cell spreading and motility through an interaction with paxillin kinase linker (PKL). J Cell Biol. 2001; 154(1):161-76. PMC: 2196859. DOI: 10.1083/jcb.200101039. View

Worthylake R, Lemoine S, Watson J, Burridge K . RhoA is required for monocyte tail retraction during transendothelial migration. J Cell Biol. 2001; 154(1):147-60. PMC: 2196864. DOI: 10.1083/jcb.200103048. View

Schuebel K, Bustelo X, Nielsen D, Song B, Barbacid M, Goldman D . Isolation and characterization of murine vav2, a member of the vav family of proto-oncogenes. Oncogene. 1996; 13(2):363-71. View

Brown M, Cary L, Jamieson J, Cooper J, Turner C . Src and FAK kinases cooperate to phosphorylate paxillin kinase linker, stimulate its focal adhesion localization, and regulate cell spreading and protrusiveness. Mol Biol Cell. 2005; 16(9):4316-28. PMC: 1196340. DOI: 10.1091/mbc.e05-02-0131. View

Nabi I . The polarization of the motile cell. J Cell Sci. 1999; 112 ( Pt 12):1803-11. DOI: 10.1242/jcs.112.12.1803. View

Machida K, Thompson C, Dierck K, Jablonowski K, Karkkainen S, Liu B . High-throughput phosphotyrosine profiling using SH2 domains. Mol Cell. 2007; 26(6):899-915. DOI: 10.1016/j.molcel.2007.05.031. View

10.

Marcoux N, Vuori K . EGF receptor mediates adhesion-dependent activation of the Rac GTPase: a role for phosphatidylinositol 3-kinase and Vav2. Oncogene. 2003; 22(38):6100-6. DOI: 10.1038/sj.onc.1206712. View

11.

Prieto-Sanchez R, Hernandez J, Garcia J, Gutierrez N, San Miguel J, Bustelo X . Overexpression of the VAV proto-oncogene product is associated with B-cell chronic lymphocytic leukaemia displaying loss on 13q. Br J Haematol. 2006; 133(6):642-5. PMC: 1950221. DOI: 10.1111/j.1365-2141.2006.06094.x. View

12.

Kim M, Nozu F, Kusama K, Imawari M . Cholecystokinin stimulates the recruitment of the Src-RhoA-phosphoinositide 3-kinase pathway by Vav-2 downstream of G(alpha13) in pancreatic acini. Biochem Biophys Res Commun. 2005; 339(1):271-6. DOI: 10.1016/j.bbrc.2005.11.014. View

13.

Nayal A, Webb D, Brown C, Schaefer E, Vicente-Manzanares M, Horwitz A . Paxillin phosphorylation at Ser273 localizes a GIT1-PIX-PAK complex and regulates adhesion and protrusion dynamics. J Cell Biol. 2006; 173(4):587-9. PMC: 2063867. DOI: 10.1083/jcb.200509075. View

14.

Tamas P, Solti Z, Buday L . Membrane-targeting is critical for the phosphorylation of Vav2 by activated EGF receptor. Cell Signal. 2001; 13(7):475-81. DOI: 10.1016/s0898-6568(01)00172-3. View

15.

Turner C . Paxillin and focal adhesion signalling. Nat Cell Biol. 2001; 2(12):E231-6. DOI: 10.1038/35046659. View

16.

Tu S, Wu W, Wang J, Cerione R . Epidermal growth factor-dependent regulation of Cdc42 is mediated by the Src tyrosine kinase. J Biol Chem. 2003; 278(49):49293-300. DOI: 10.1074/jbc.M307021200. View

17.

Billadeau D . Cell growth and metastasis in pancreatic cancer: is Vav the Rho'd to activation?. Int J Gastrointest Cancer. 2003; 31(1-3):5-13. DOI: 10.1385/IJGC:31:1-3:5. View

18.

Booden M, Campbell S, Der C . Critical but distinct roles for the pleckstrin homology and cysteine-rich domains as positive modulators of Vav2 signaling and transformation. Mol Cell Biol. 2002; 22(8):2487-97. PMC: 133724. DOI: 10.1128/MCB.22.8.2487-2497.2002. View

19.

Liu B, Burridge K . Vav2 activates Rac1, Cdc42, and RhoA downstream from growth factor receptors but not beta1 integrins. Mol Cell Biol. 2000; 20(19):7160-9. PMC: 86269. DOI: 10.1128/MCB.20.19.7160-7169.2000. View

20.

Wormer D, Deakin N, Turner C . CdGAP regulates cell migration and adhesion dynamics in two-and three-dimensional matrix environments. Cytoskeleton (Hoboken). 2012; 69(9):644-58. PMC: 4474383. DOI: 10.1002/cm.21057. View