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PMID: 9658172 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Serine and threonine phosphorylation of the paxillin LIM domains regulates paxillin focal adhesion localization and cell adhesion to fibronectin.

Molecular biology of the cell ·Vol. 9 ·No. 7 ·1998-07-00 ·Pages 1803-16

Brown MC, Perrotta JA, Turner CE

Abstract

We have previously shown that the LIM domains of paxillin operate as the focal adhesion (FA)-targeting motif of this protein. In the current study, we have identified the capacity of paxillin LIM2 and LIM3 to serve as binding sites for, and substrates of serine/threonine kinases. The activities of the LIM2- and LIM3-associated kinases were stimulated after adhesion of CHO.K1 cells to fibronectin; consequently, a role for LIM domain phosphorylation in regulating the subcellular localization of paxillin after adhesion to fibronectin was investigated. An avian paxillin-CHO.K1 model system was used to explore the role of paxillin phosphorylation in paxillin localization to FAs. We found that mutations of paxillin that mimicked LIM domain phosphorylation accelerated fibronectin-induced localization of paxillin to focal contacts. Further, blocking phosphorylation of the LIM domains reduced cell adhesion to fibronectin, whereas constitutive LIM domain phosphorylation significantly increased the capacity of cells to adhere to fibronectin. The potentiation of FA targeting and cell adhesion to fibronectin was specific to LIM domain phosphorylation as mutation of the amino-terminal tyrosine and serine residues of paxillin that are phosphorylated in response to fibronectin adhesion had no effect on the rate of FA localization or cell adhesion. This represents the first demonstration of the regulation of protein localization through LIM domain phosphorylation and suggests a novel mechanism of regulating LIM domain function. Additionally, these results provide the first evidence that paxillin contributes to "inside-out" integrin-mediated signal transduction.

MeSH Terms
Amino Acid Sequence Animals CHO Cells Cell Adhesion Clone Cells Cricetinae Cytoskeletal Proteins/metabolism,physiology DNA Mutational Analysis Fibroblasts/metabolism Fibronectins/metabolism Molecular Sequence Data Mutagenesis, Site-Directed Paxillin Peptide Fragments/metabolism Phosphoproteins/metabolism,physiology Phosphorylation Recombinant Fusion Proteins/genetics,metabolism Serine/metabolism Threonine/metabolism
Chemicals
Cytoskeletal Proteins Fibronectins Paxillin Peptide Fragments Phosphoproteins Recombinant Fusion Proteins Threonine Serine
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Brown M C
Department of Anatomy and Cell Biology, Program in Cell and Molecular Biology, State University of New York Health Science Center at Syracuse, Syracuse, New York 13210, USA.
Perrotta J A
Turner C E
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Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.9.7.1803
Published
1998-07-00
Pages
1803-16
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC25420
Subset
IM
Grants
NIGMS NIH HHS · R01 GM047607 · United States
NIGMS NIH HHS · GM-47607 · United States
External Links
PubMed source DOI: 10.1091/mbc.9.7.1803 PMC Full Text View journal details
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