Tyrosine phosphorylation of β-dystroglycan at its WW domain binding motif, PPxY, recruits SH2 domain containing proteins

F. Sotgia, H. Lee, M. T. Bedford, T. Petrucci, M. Sudol, M. P. Lisanti

Research output: Contribution to journalArticle

78 Citations (Scopus)

Abstract

β-Dystroglycan is a ubiquitously expressed integral membrane protein that undergoes tyrosine phosphorylation in an adhesion-dependent manner. However, it remains unknown whether tyrosine-phosphorylated β-dystroglycan interacts with SH2 domain containing proteins. Here, we show that the tyrosine phosphorylation of β-dystroglycan is constitutively elevated in v-Src transformed cells. We next reconstituted this phosphorylation event in vivo by transiently coexpressing wild-type c-Src with a fusion protein containing full-length β-dystroglycan. Our results demonstrate that Src-induced tyrosine phosphorylation of β-dystroglycan is strictly dependent on the presence of a PPxY motif at its extreme C-terminus. In the nonphosphorylated state, this PPxY motif is normally recognized as a ligand by the WW domain; phosphorylation at this site blocks the binding of certain WW domain containing proteins. Using a GST fusion protein carrying the cytoplasmic tail of β-dystroglycan, we identified five SH2 domain containing proteins that interact with β-dystroglycan in a phosphorylation-dependent manner, including c-Src, Fyn, Csk, NCK, and SHC. We localized this binding activity to the PPxY motif by employing a panel of β-dystroglycan-derived phosphopeptides. In addition, tyrosine phosphorylation of β-dystroglycan in vivo resulted in the coimmunoprecipitation of the same SH2 domain containing proteins, and this binding event required the β-dystroglycan C-terminal PPxY motif. We discuss the possibility that tyrosine phosphorylation of the PPxY motif within β-dystroglycan may act as a regulatory switch to inhibit the binding of certain WW domain containing proteins, while recruiting SH2 domain containing proteins.

Original languageEnglish
Pages (from-to)14585-14592
Number of pages8
JournalBiochemistry
Volume40
Issue number48
DOIs
Publication statusPublished - 2001 Dec 4

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Dystroglycans
Phosphorylation
src Homology Domains
Tyrosine
Proteins
Fusion reactions
Phosphopeptides
Protein Binding
Membrane Proteins

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Sotgia, F. ; Lee, H. ; Bedford, M. T. ; Petrucci, T. ; Sudol, M. ; Lisanti, M. P. / Tyrosine phosphorylation of β-dystroglycan at its WW domain binding motif, PPxY, recruits SH2 domain containing proteins. In: Biochemistry. 2001 ; Vol. 40, No. 48. pp. 14585-14592.
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Tyrosine phosphorylation of β-dystroglycan at its WW domain binding motif, PPxY, recruits SH2 domain containing proteins. / Sotgia, F.; Lee, H.; Bedford, M. T.; Petrucci, T.; Sudol, M.; Lisanti, M. P.

In: Biochemistry, Vol. 40, No. 48, 04.12.2001, p. 14585-14592.

Research output: Contribution to journalArticle

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AU - Sotgia, F.

AU - Lee, H.

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AU - Lisanti, M. P.

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