Individual Cas phosphorylation sites are dispensable for processive phosphorylation by Src and anchorage-independent cell growth

Parag Patwardhan, Yongquan Shen, Gary S. Goldberg, W. Todd Miller

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

Cas is a multidomain signaling protein that resides in focal adhesions. Cas possesses a large central substrate domain containing 15 repeats of the sequence YXXP, which are phosphorylated by Src. The phosphorylation sites are essential for the roles of Cas in cell migration and in regulation of the actin cytoskeleton. We showed previously that Src catalyzes the multisite phosphorylation of Cas via a processive mechanism. In this study,wecreated mutant forms of Cas to identify the determinants for processive phosphorylation. Mutants containing single or multiple YXXP mutations were phosphorylated processively by Src, suggesting that individual sites are dispensable. The results also suggest that there is no defined order to the Cas phosphorylation events. We also studied the effects of these mutations by reintroducing Cas into Cas-deficient fibroblasts. Mutants lacking some or all YXXP sites augment the ability of Src to promote anchorage-independent growth. On the other hand, deletion of YXXP sites compromises the ability of Cas to promote tumor cell migration.

Original languageEnglish (US)
Pages (from-to)20689-20697
Number of pages9
JournalJournal of Biological Chemistry
Volume281
Issue number30
DOIs
StatePublished - Jul 28 2006

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Phosphorylation
Cell growth
Growth
Cell Movement
Mutation
Focal Adhesions
Fibroblasts
Actin Cytoskeleton
Actins
Tumors
Adhesion
Cells
Substrates
Neoplasms
Proteins

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

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abstract = "Cas is a multidomain signaling protein that resides in focal adhesions. Cas possesses a large central substrate domain containing 15 repeats of the sequence YXXP, which are phosphorylated by Src. The phosphorylation sites are essential for the roles of Cas in cell migration and in regulation of the actin cytoskeleton. We showed previously that Src catalyzes the multisite phosphorylation of Cas via a processive mechanism. In this study,wecreated mutant forms of Cas to identify the determinants for processive phosphorylation. Mutants containing single or multiple YXXP mutations were phosphorylated processively by Src, suggesting that individual sites are dispensable. The results also suggest that there is no defined order to the Cas phosphorylation events. We also studied the effects of these mutations by reintroducing Cas into Cas-deficient fibroblasts. Mutants lacking some or all YXXP sites augment the ability of Src to promote anchorage-independent growth. On the other hand, deletion of YXXP sites compromises the ability of Cas to promote tumor cell migration.",
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Individual Cas phosphorylation sites are dispensable for processive phosphorylation by Src and anchorage-independent cell growth. / Patwardhan, Parag; Shen, Yongquan; Goldberg, Gary S.; Miller, W. Todd.

In: Journal of Biological Chemistry, Vol. 281, No. 30, 28.07.2006, p. 20689-20697.

Research output: Contribution to journalArticle

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