PTP-PEST targets a novel tyrosine site in p120 catenin to control epithelial cell motility and Rho GTPase activity.

Abstract

Tyrosine phosphorylation is implicated in regulating the adherens junction protein, p120 catenin (p120), however, the mechanisms are not well defined. Here, we show, using substrate trapping, that p120 is a direct target of the protein tyrosine phosphatase, PTP-PEST, in epithelial cells. Stable shRNA knockdown of PTP-PEST in colon carcinoma cells results in an increased cytosolic pool of p120 concomitant with its enhanced tyrosine phosphorylation and decreased association with E-cadherin. Consistent with this, PTP-PEST knockdown cells exhibit increased motility, enhanced Rac1 and decreased RhoA activity on a collagen substrate. Furthermore, p120 localization is enhanced at actin-rich protrusions and lamellipodia and has an increased association with the guanine nucleotide exchange factor, VAV2, and cortactin. Exchange factor activity of VAV2 is enhanced by PTP-PEST knockdown whereas overexpression of a VAV2 C-terminal domain or DH domain mutant blocks cell motility. Analysis of point mutations identified tyrosine 335 in the N-terminal domain of p120 as the site of PTP-PEST dephosphorylation. A Y335F mutant of p120 failed to induce the 'p120 phenotype', interact with VAV2, stimulate cell motility or activate Rac1. Together, these data suggest that PTP-PEST affects epithelial cell motility by controlling the distribution and phosphorylation of p120 and its availability to control Rho GTPase activity.

• Authors: Rosario Espejo, Yowjiun Jeng, Adriana Paulucci-Holthauzen, William Rengifo-Cam, Krysta Honkus, Panos Z Anastasiadis, Sarita K Sastry
• Journal: Journal of cell science (J Cell Sci) Vol. 127 Issue Pt 3 Pg. 497-508 (Feb 01 2014) ISSN: 1477-9137 [Electronic] England
• PMID: 24284071 (Publication Type: Journal Article, Research Support, N.I.H., Extramural)

Chemical References

• p120 GTPase Activating Protein
• Tyrosine
• PTPN12 protein, human
• Protein Tyrosine Phosphatase, Non-Receptor Type 12
• rho GTP-Binding Proteins

Topics

• Cell Line, Tumor
• Cell Movement (genetics)
• Colonic Neoplasms (genetics, metabolism, pathology)
• Epithelial Cells
• Humans
• Mutation
• Phosphorylation (genetics)
• Protein Tyrosine Phosphatase, Non-Receptor Type 12 (genetics, metabolism)
• Tyrosine (genetics)
• p120 GTPase Activating Protein (genetics, metabolism)
• rho GTP-Binding Proteins (genetics, metabolism)