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PMID: 11149924 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Targeting Pyk2 to beta 1-integrin-containing focal contacts rescues fibronectin-stimulated signaling and haptotactic motility defects of focal adhesion kinase-null cells.

The Journal of cell biology ·Vol. 152 ·No. 1 ·2001-01-08 ·Pages 97-110

Klingbeil CK, Hauck CR, Hsia DA, Jones KC, Reider SR, Schlaepfer DD

Abstract

Focal adhesion kinase-null (FAK(-/-) fibroblasts exhibit morphological and motility defects that are reversed by focal adhesion kinase (FAK) reexpression. The FAK-related kinase, proline-rich tyrosine kinase 2 (Pyk2), is expressed in FAK(-/-) cells, yet it exhibits a perinuclear distribution and does not functionally substitute for FAK. Chimeric Pyk2/FAK proteins were created and expressed in FAK(-/-) cells to determine the impact of Pyk2 localization to focal contacts. Whereas an FAK/Pyk2 COOH-terminal (CT) domain chimera was perinuclear distributed, stable expression of a Pyk2 chimera with the FAK-CT domain (Pyk2/FAK-CT) localized to focal contact sites and enhanced fibronectin (FN)-stimulated haptotactic cell migration equal to FAK-reconstituted cells. Disruption of paxillin binding to the FAK-CT domain (S-1034) inhibited Pyk2/FAK-CT localization to focal contacts and its capacity to promote cell motility. Paxillin binding to the FAK-CT was necessary but not sufficient to mediate the indirect association of FAK or Pyk2/FAK-CT with a beta 1-integrin-containing complex. Both FAK and Pyk2/FAK-CT but not Pyk2/FAK-CT S-1034 reconstituted FAK(-/-) cells, exhibit elevated FN-stimulated extracellular signal-regulated kinase 2 (ERK2) and c-Jun NH(2)-terminal kinase (JNK) kinase activation. FN-stimulated FAK or Pyk2/FAK-CT activation enhanced both the extent and duration of FN-stimulated ERK2 activity which was necessary for cell motility. Transient overexpression of the FAK-CT but not FAK-CT S-1034 domain inhibited both FN-stimulated ERK2 and JNK activation as well as FN-stimulated motility of Pyk2/FAK-CT reconstituted cells. These gain-of-function studies show that the NH(2)-terminal and kinase domains of Pyk2 can functionally substitute for FAK in promoting FN-stimulated signaling and motility events when localized to beta-integrin-containing focal contact sites via interactions mediated by the FAK-CT domain.

MeSH Terms
Animals Cell Movement/physiology Cells, Cultured Cytoskeletal Proteins/metabolism Enzyme Activation Fibroblasts/cytology,metabolism,physiology Fibronectins/metabolism Focal Adhesion Kinase 1 Focal Adhesion Kinase 2 Focal Adhesion Protein-Tyrosine Kinases Gene Expression Humans Integrin beta1/metabolism JNK Mitogen-Activated Protein Kinases Mice Mice, Knockout Mitogen-Activated Protein Kinase 1/metabolism Mitogen-Activated Protein Kinases/metabolism Paxillin Phosphoproteins/metabolism Protein-Tyrosine Kinases/genetics,metabolism Recombinant Fusion Proteins/genetics,metabolism Signal Transduction
Chemicals
Cytoskeletal Proteins Fibronectins Integrin beta1 PXN protein, human Paxillin Phosphoproteins Pxn protein, mouse Recombinant Fusion Proteins Protein-Tyrosine Kinases Focal Adhesion Kinase 1 Focal Adhesion Kinase 2 Focal Adhesion Protein-Tyrosine Kinases PTK2 protein, human Ptk2 protein, mouse Ptk2b protein, mouse JNK Mitogen-Activated Protein Kinases Mitogen-Activated Protein Kinase 1 Mitogen-Activated Protein Kinases
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Klingbeil C K
Department of Immunology, The Scripps Research Institute, La Jolla, California 92037, USA.
Hauck C R
Hsia D A
Jones K C
Reider S R
Schlaepfer D D
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.152.1.97
Published
2001-01-08
Pages
97-110
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2193658
Subset
IM
Grants
NCI NIH HHS · CA75240 · United States
External Links
PubMed source DOI: 10.1083/jcb.152.1.97 PMC Full Text View journal details
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