F. Sotgia et al., Tyrosine phosphorylation of beta-dystroglycan at its WW domain binding motif, PPxY, recruits SH2 domain containing proteins, BIOCHEM, 40(48), 2001, pp. 14585-14592
beta -Dystroglycan is a ubiquitously expressed integral membrane protein th
at undergoes tyrosine phosphorylation in an adhesion-dependent manner. Howe
ver, it remains unknown whether tyrosine-phosphorylated beta -dystroglycan
interacts with SH2 domain containing proteins. Here, we show that the tyros
ine phosphorylation of l3-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 containi
ng full-length beta -dystroglycan. Our results demonstrate that Src-induced
tyrosine phosphorylation of beta -dystroglycan is strictly dependent on th
e presence of a PPxY motif at its extreme C-terminus. In the nonphosphoryla
ted state, this PPxY motif is normally recognized as a ligand by the WW dom
ain; phosphorylation at this site blocks the binding of certain WW domain c
ontaining proteins. Using a GST fusion protein carrying the cytoplasmic tai
l of beta -dystroglycan, we identified five SH2 domain containing proteins
that interact with beta -dystroglycan in a phosphorylation-dependent manner
, including c-Src, Fyn, Csk, NCK, and SHC. We localized this binding activi
ty to the PPxY motif by employing a panel of beta -dystroglycan-derived pho
sphopeptides. In addition, tyrosine phosphorylation of beta -dystroglycan i
n vivo resulted in the coimmunoprecipitation of the same SH2 domain contain
ing proteins, and this binding event required the -dystroglycan C-terminal
PPxY motif. We discuss the possibility that tyrosine phosphorylation of the
PPxY motif within beta -dystroglycan may act as a regulatory switch to inh
ibit the binding of certain WW domain containing proteins, while recruiting
SH2 domain containing proteins.