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PMID: 10982414 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Paxillin binding is not the sole determinant of focal adhesion localization or dominant-negative activity of focal adhesion kinase/focal adhesion kinase-related nonkinase.

Molecular biology of the cell ·Vol. 11 ·No. 9 ·2000-09-00 ·Pages 3247-63

Cooley MA, Broome JM, Ohngemach C, Romer LH, Schaller MD

Abstract

The carboxy-terminal 150 residues of the focal adhesion kinase (FAK) comprise the focal adhesion-targeting sequence, which is responsible for its subcellular localization. The mechanism of focal adhesion targeting has not been fully elucidated. We describe a mutational analysis of the focal adhesion-targeting sequence of FAK to further examine the mechanism of focal adhesion targeting and explore additional functions encoded by the carboxy-terminus of FAK. The results demonstrate that paxillin binding is dispensable for focal adhesion targeting of FAK. Cell adhesion-dependent tyrosine phosphorylation strictly correlated with the ability of mutants to target to focal adhesions. Focal adhesion targeting was also a requirement for maximal FAK-dependent tyrosine phosphorylation of paxillin and FAK-related nonkinase (FRNK)-dependent inhibition of endogenous FAK function. However, there were additional requirements for these latter functions because we identified mutants that target to focal adhesions, yet are defective for the induction of paxillin phosphorylation or the dominant-negative function of FRNK. Furthermore, the paxillin-binding activity of FRNK mutants did not correlate with their ability to inhibit FAK, suggesting that FRNK has other targets in addition to paxillin.

MeSH Terms
3T3 Cells Amino Acid Sequence Animals Binding Sites Cells, Cultured Chick Embryo Cytoskeletal Proteins/chemistry,metabolism Focal Adhesion Kinase 1 Focal Adhesion Protein-Tyrosine Kinases Genetic Variation Mice Molecular Sequence Data Mutagenesis, Site-Directed Paxillin Phosphoproteins/chemistry,metabolism Protein-Tyrosine Kinases/chemistry,genetics,metabolism Recombinant Fusion Proteins/metabolism Sequence Alignment Sequence Homology, Amino Acid Subcellular Fractions/metabolism,ultrastructure Transfection
Chemicals
Cytoskeletal Proteins Paxillin Phosphoproteins Pxn protein, mouse Recombinant Fusion Proteins FAK-related nonkinase Protein-Tyrosine Kinases Focal Adhesion Kinase 1 Focal Adhesion Protein-Tyrosine Kinases Ptk2 protein, mouse
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Cooley M A
Departments of Cell Biology & Anatomy, University of North Carolina, Chapel Hill, North Carolina 27599, USA.
Broome J M
Ohngemach C
Romer L H
Schaller M D
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Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.11.9.3247
Published
2000-09-00
Pages
3247-63
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC14989
Subset
IM
External Links
PubMed source DOI: 10.1091/mbc.11.9.3247 PMC Full Text View journal details
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