RGD Reference Report - p130cas but not paxillin is essential for Caco-2 intestinal epithelial cell spreading and migration on collagen IV. - Rat Genome Database

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p130cas but not paxillin is essential for Caco-2 intestinal epithelial cell spreading and migration on collagen IV.

Authors: Sanders, MA  Basson, MD 
Citation: Sanders MA and Basson MD, J Biol Chem. 2005 Jun 24;280(25):23516-22. Epub 2005 Apr 6.
RGD ID: 7488898
Pubmed: PMID:15817476   (View Abstract at PubMed)
DOI: DOI:10.1074/jbc.M413165200   (Journal Full-text)

We have previously observed that collagen IV regulates Caco-2 intestinal epithelial cell spreading and migration via Src kinase and stimulates Src-dependent tyrosine phosphorylation of p130cas. We observed that collagen IV also stimulated Src-dependent phosphorylation of both paxillin Tyr31 and paxillin Tyr118. Caco-2 transfection with paxillin or p130cas siRNAs inhibited expression of these proteins by more than 90% for at least 5 days after transfection. Although p130cas siRNA inhibited cell spreading on collagen IV by 33%, three different paxillin siRNAs did not inhibit cell spreading. p130cas siRNA did not affect Src Tyr416 or Src Tyr527 phosphorylation, FAK Tyr397 phosphorylation, or Src-dependent phosphorylation of FAK Tyr925, suggesting that p130cas did not inhibit cell spreading by altering FAK or Src activity. Rat p130cas expression after siRNA knock-out of endogenous human p130cas in Caco-2 cells reduced cell spreading inhibition by 71%. In contrast, expression of rat p130cas from which the Src-phosphorylated substrate domain was deleted did not rescue siRNA inhibition of cell spreading. Combined treatment with siRNAs to Crk and CrkL, which bind to the p130cas substrate domain, inhibited cell spreading by 54%. Both p130cas siRNA and the combined Crk/CrkL siRNAs strongly inhibited (52 and 46% inhibition, respectively) Caco-2 sheet migration on collagen IV and noticeably inhibited lamellipodial extension, whereas paxillin siRNA only inhibited migration by 18% and did not noticeably affect lamellipodial extension. These results suggest that Src may regulate Caco-2 migration on collagen IV via both p130cas and paxillin but that Src phosphorylation of p130cas is more important for this process.

Gene Ontology Annotations    Click to see Annotation Detail View

Biological Process
TermQualifierEvidenceWithReferenceNotesSourceOriginal Reference(s)
negative regulation of substrate adhesion-dependent cell spreading  IMP 7488898 RGD 

Objects Annotated

Genes (Rattus norvegicus)
Bcar1  (BCAR1 scaffold protein, Cas family member)


Additional Information