PTK6 Localized at the Plasma Membrane Promotes Cell Proliferation and MigratiOn Through Phosphorylation of Eps8. Issue 9 (15th May 2017)

• Record Type: Journal Article
• Authors: Shin, Won‐Sik; Shim, Hyun Jae; Lee, Young Hun; Pyo, Minju; Park, Jun Sang; Ahn, So Yun; Lee, Seung‐Taek
• Is Part Of: Journal of cellular biochemistry. Volume 118:Issue 9(2017)
• Page Start: 2887
• Page End: 2895
• Publication Date: 2017-05-15
• Subjects: EPS8 -- PTK6 -- PROLIFERATION -- MIGRATION -- TUMORIGENESIS; Cytochemistry -- Periodicals; 572
• DOI: 10.1002/jcb.25939
• Languages: English
• ISSNs: 0730-2312

Abstract

ABSTRACT: Protein tyrosine kinase 6 (PTK6; also known as Brk) is closely related to the Src family kinases, but lacks a membrane‐targeting myristoylation signal. Sublocalization of PTK6 at the plasma membrane enhances its oncogenic potential. To understand the mechanism(s) underlying the oncogenic property of plasma­­–membrane‐associated PTK6, proteins phosphorylated by membrane‐targeted myristoylated PTK6 (Myr‐PTK6) were enriched and analyzed using a proteomics approach. Eps8 which was identified by this method is phosphorylated by Myr‐PTK6 in HEK293 cells. Mouse Eps8 expressed in HEK293 cells is phosphorylated by Myr‐PTK6 at residues Tyr497, Tyr524, and Tyr534. Compared to wild‐type Eps8 (Eps8 WT), the phosphorylation‐defective 3YF mutant (Eps8 3YF) reverts the increased proliferation, migration, and phosphorylation of ERK and FAK mediated by Eps8 WT in HEK293 cells overexpressing PTK6. PTK6 knockdown in T‐47D breast cancer cells decreased EGF‐induced phosphorylation of Eps8. Endogenous PTK6 phosphorylates ectopically expressed Eps8 WT, but not Eps8 3YF mutant, in EGF‐stimulated T‐47D cells. The EGF‐induced Eps8 phosphorylation enhances activation of ERK and FAK, cell adhesion, and anchorage‐independent colony formation in T‐47D cells, but not in the PTK6‐knokdown T‐47D cells. These results indicate that plasma–membrane‐associated PTK6 phosphorylates Eps8, which promotes cell proliferation, adhesion, and migration and, thus, tumorigenesis. J. Cell. Biochem. 118: 2887–2895, 2017. © 2017 Wiley Periodicals, Inc. Abstract : EGFR pathway substrate 8 (Eps8) was identified as a novel substrate of plasma­­–membrane‐associated PTK6. Phosphorylation‐defective mutant of Eps8 poorly promotes proliferation, migration, and phosphorylation of ERK and FAK. Knockdown of PTK6 significantly decreases EGF‐stimulated Eps8 phosphorylation, adhesion, and anchorage‐independent colony formation in breast cancer cells. Identification of Eps8 as a substrate of activated PTK6 is important for controlling PTK6‐positive diseases such as breast cancers.