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A yeast two‐hybrid study of human p97/Gab2 interactions with its SH2 domain‐containing binding partners
Author(s) -
Crouin Catherine,
Arnaud Mary,
Gesbert Franck,
Camonis Jacques,
Bertoglio Jacques
Publication year - 2001
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(01)02373-0
Subject(s) - sh2 domain , tyrosine , chemistry , protein tyrosine phosphatase , protein subunit , yeast , microbiology and biotechnology , proto oncogene tyrosine protein kinase src , tyrosine kinase , signal transducing adaptor protein , lyn , biochemistry , protein–protein interaction , phosphotyrosine binding domain , signal transduction , biology , gene
p97/Gab2 is a recently characterized member of a large family of scaffold proteins that play essential roles in signal transduction. Gab2 becomes tyrosine‐phosphorylated in response to a variety of growth factors and forms multimolecular complexes with SH2 domain‐containing signaling molecules such as the p85‐regulatory subunit of the phosphoinositide‐3‐kinase (p85‐PI3K), the tyrosine phosphatase SHP‐2 and the adapter protein CrkL. To characterize the interactions between Gab2 and its SH2‐containing binding partners, we designed a modified yeast two‐hybrid system in which the Lyn tyrosine kinase is expressed in a regulated manner in yeast. Using this assay, we demonstrated that p97/Gab2 specifically interacts with the SH2 domains of PI3K, SHP‐2 and CrkL. Interaction with p85‐PI3K is mediated by tyrosine residues Y 452 , Y 476 and Y 584 of Gab2, while interaction with SHP‐2 depends exclusively on tyrosine Y 614 . CrkL interaction is mediated by its SH2 domain recognizing Y 266 and Y 293 , despite the latter being in a non‐consensus (YTFK) environment.