Mutational analysis of the SRC homology 2 domain protein-tyrosine phosphatase Corkscrew.
The SRC homology 2 (SH2) domain protein-tyrosine phosphatase, Corkscrew (CSW) is required for signaling by receptor tyrosine kinases, including the Sevenless receptor tyrosine kinase (SEV), which directs Drosophila R7 photoreceptor cell development. To investigate the role of the different domains of CSW, we constructed domain-specific csw mutations and assayed their effects on CSW function. Our results indicate that CSW SH2 domain function is essential, but either CSW SH2 domain can fulfill this requirement. We also found that CSW and activated SEV are associated in vivo in a manner that does not require either CSW SH2 domain function or tyrosine phosphorylation of SEV. In contrast, the interaction between CSW and Daughter of Sevenless, a CSW substrate, is dependent on SH2 domain function. These results suggest that the role of the CSW SH2 domains during SEV signaling is to bind Daughter of Sevenless rather than activated SEV. We also found that although CSW protein-tyrosine phosphatase activity is required for full CSW function, a catalytically inactive CSW is capable of providing partial function. In addition, we found that deletion of either the CSW protein- tyrosine phosphatase insert or the entire CSW carboxyl terminus, which includes a conserved DRK/GRB2 SH2 domain binding sequence, does not abolish CSW function.
Allard
J D
JD
Department of Biological Sciences, Stanford University, Stanford, California 94305-5020, USA.
Herbst
R
R
Carroll
P M
PM
Simon
M A
MA
D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D001483:Base Sequence; D002384:Catalysis; D002460:Cell Line; D017931:DNA Primers; D004330:Drosophila; D029721:Drosophila Proteins; D005260:Female; D051380:GRB2 Adaptor Protein; D008297:Male; D016296:Mutagenesis; D010766:Phosphorylation; D017956:Photoreceptor Cells, Invertebrate / Q000378:metabolism; D017027:Protein Tyrosine Phosphatases / Q000235:genetics / Q000378:metabolism*; D054558:Protein Tyrosine Phosphatases, Non-Receptor; D011506:Proteins / Q000378:metabolism; D015398:Signal Transduction; D013379:Substrate Specificity; D018909:src Homology Domains
D048868:Adaptor Proteins, Signal Transducing; D017931:DNA Primers; D029721:Drosophila Proteins; D051380:GRB2 Adaptor Protein; D011506:Proteins; C098692:Csw protein, Drosophila; D017027:Protein Tyrosine Phosphatases; D054558:Protein Tyrosine Phosphatases, Non-Receptor