Enhancement of tyrosyl phosphorylation and protein expression of eps8 by v-Src.
Two eps8 isoforms, p97eps8 and p68eps8, were previously identified as substrates for receptor tyrosine kinases. Analysis of eps8 phosphotyrosine content in v-Src transformed cells (IV5) revealed that both isoforms were highly tyrosyl phosphorylated and their readiness to be phosphorylated by Src in vitro further indicated that they were putative Src substrates as well. Indeed, the enhancement of tyrosyl phosphorylation of p97eps8 detected in cells coexpressing both p97eps8 and active Src relative to that in cells expressing p97eps8 alone supported our hypothesis. The existence of common phosphotryptic peptides between in vitro 32P-labeled p97eps8 and p68eps8 indicated that these two proteins shared the same Src-mediated sites. Further in vitro binding assays demonstrated that p68eps8 was the major eps8 isoforms that could be precipitated by bacterial fusion protein containing Src SH3. Interestingly, both p68eps8 and p97eps8 were preferentially expressed in v-Src transformed cells and the presence of p68eps8 appeared to depend on Src. Since p97eps8 has been implicated in mitogenesis and tumorigenesis, its readiness to be phosphorylated and induced by v-Src might attribute to v-Src-mediated transformation.
Institute of Biochemistry, Chung Shan Medical and Dental College, No. 113, Section 2, Ta-Ching Street, Taichung 402, Taiwan.
M C
MC
Maa
J R
JR
Lai
R W
RW
Lin
T H
TH
Leu
D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D002461:Cell Line, Transformed; D002478:Cells, Cultured; D002642:Chick Embryo; D003001:Cloning, Molecular; D003598:Cytoskeletal Proteins; D005786:Gene Expression Regulation / Q000187:drug effects; D051379:Mice; D015688:Oncogene Protein pp60(v-src) / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology; D010766:Phosphorylation / Q000187:drug effects; D014176:Protein Biosynthesis; D011506:Proteins / Q000737:chemistry / Q000235:genetics; D014443:Tyrosine / Q000737:chemistry*; D018909:src Homology Domains
D048868:Adaptor Proteins, Signal Transducing; D003598:Cytoskeletal Proteins; C083430:Eps8 protein, mouse; D011506:Proteins; D014443:Tyrosine; D015688:Oncogene Protein pp60(v-src)