Cortactin Phosphorylated by ERK1/2 Localizes to Sites of Dynamic Actin Regulation and Is Required for Carcinoma Lamellipodia Persistence

Authors

Laura C. Kelley, West Virginia University
Karen E. Hayes, West Virginia University
Amanda G. Ammer, West Virginia University
Karen H. Martin, West Virginia University
Scott A. Weed, West Virginia University

Author ORCID Identifier

https://orcid.org/0000-0003-2706-3610

N/A

https://orcid.org/0000-0002-0790-8033

https://orcid.org/0000-0001-8916-4092

https://orcid.org/0000-0001-7033-9999

Document Type

Article

Publication Date

2010

College/Unit

School of Medicine

Department/Program/Center

Neurology

Abstract

Background

Tumor cell motility and invasion is governed by dynamic regulation of the cortical actin cytoskeleton. The actin-binding protein cortactin is commonly upregulated in multiple cancer types and is associated with increased cell migration. Cortactin regulates actin nucleation through the actin related protein (Arp)2/3 complex and stabilizes the cortical actin cytoskeleton. Cortactin is regulated by multiple phosphorylation events, including phosphorylation of S405 and S418 by extracellular regulated kinases (ERK)1/2. ERK1/2 phosphorylation of cortactin has emerged as an important positive regulatory modification, enabling cortactin to bind and activate the Arp2/3 regulator neuronal Wiskott-Aldrich syndrome protein (N-WASp), promoting actin polymerization and enhancing tumor cell movement.

Methodology/Principal Findings

In this report we have developed phosphorylation-specific antibodies against phosphorylated cortactin S405 and S418 to analyze the subcellular localization of this cortactin form in tumor cells and patient samples by microscopy. We evaluated the interplay between cortactin S405 and S418 phosphorylation with cortactin tyrosine phosphorylation in regulating cortactin conformational forms by Western blotting. Cortactin is simultaneously phosphorylated at S405/418 and Y421 in tumor cells, and through the use of point mutant constructs we determined that serine and tyrosine phosphorylation events lack any co-dependency. Expression of S405/418 phosphorylation-null constructs impaired carcinoma motility and adhesion, and also inhibited lamellipodia persistence monitored by live cell imaging.

Conclusions/Significance

Cortactin phosphorylated at S405/418 is localized to sites of dynamic actin assembly in tumor cells. Concurrent phosphorylation of cortactin by ERK1/2 and tyrosine kinases enables cells with the ability to regulate actin dynamics through N-WASp and other effector proteins by synchronizing upstream regulatory pathways, confirming cortactin as an important integration point in actin-based signal transduction. Reduced lamellipodia persistence in cells with S405/418A expression identifies an essential motility-based process reliant on ERK1/2 signaling, providing additional understanding as to how this pathway impacts tumor cell migration.

Digital Commons Citation

Kelley, Laura C.; Hayes, Karen E.; Ammer, Amanda G.; Martin, Karen H.; and Weed, Scott A., "Cortactin Phosphorylated by ERK1/2 Localizes to Sites of Dynamic Actin Regulation and Is Required for Carcinoma Lamellipodia Persistence" (2010). Faculty & Staff Scholarship. 2776.
https://researchrepository.wvu.edu/faculty_publications/2776

Source Citation

Kelley LC, Hayes KE, Ammer AG, Martin KH, Weed SA (2010) Cortactin Phosphorylated by ERK1/2 Localizes to Sites of Dynamic Actin Regulation and Is Required for Carcinoma Lamellipodia Persistence. PLoS ONE 5(11): e13847. https://doi.org/10.1371/journal.pone.0013847

Comments

© 2010 Kelley et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.