Stern M J, Marengere L E, Daly R J, Lowenstein E J, Kokel M, Batzer A, Olivier P, Pawson T, Schlessinger J
Department of Genetics, Yale University School of Medicine, New Haven, Connecticut 06520-8005.
Mol Biol Cell. 1993 Nov;4(11):1175-88. doi: 10.1091/mbc.4.11.1175.
Mutations in the Caenorhabditis elegans gene sem-5 affect cell signaling processes involved in guiding a class of cell migrations and inducing vulval cell fates. The sem-5 sequence encodes a protein comprised almost exclusively of SH2 and SH3 domains (SH, src homology region) that are found together in many signaling proteins and nonreceptor tyrosine kinases. A human protein, GRB2, was identified by its ability to associate with the activated human epidermal growth factor receptor (hEGFR). The GRB2 and Sem-5 proteins share an identical architecture of their SH2 and SH3 domains and 58% amino acid sequence identity. Here we demonstrate that GRB2 and a Drosophila sem-5-like gene Drk can specifically rescue sem-5 mutants. We also show that Sem-5, like GRB2, can bind to the activated hEGFR in vitro. We further correlate the abilities of several mutant variants of GRB2 and Sem-5 to bind to the hEGFR in vitro with their abilities to functionally replace sem-5 in vivo. These data indicate that GRB2 and Drk are functional homologues of Sem-5 and demonstrate the high degree of conservation of both structure and function between signaling systems throughout evolution.
秀丽隐杆线虫基因sem-5的突变会影响一类细胞迁移导向及诱导外阴细胞命运所涉及的细胞信号传导过程。sem-5序列编码一种几乎完全由SH2和SH3结构域(SH,src同源区域)组成的蛋白质,这些结构域在许多信号蛋白和非受体酪氨酸激酶中共同存在。一种人类蛋白质GRB2因其与活化的人类表皮生长因子受体(hEGFR)结合的能力而被鉴定出来。GRB2和Sem-5蛋白的SH2和SH3结构域具有相同的结构,氨基酸序列同一性为58%。在这里,我们证明GRB2和一个果蝇sem-5样基因Drk可以特异性地挽救sem-5突变体。我们还表明,Sem-5与GRB2一样,在体外可以结合活化的hEGFR。我们进一步将GRB2和Sem-5的几种突变变体在体外结合hEGFR的能力与其在体内功能替代sem-5的能力相关联。这些数据表明GRB2和Drk是Sem-5的功能同源物,并证明了整个进化过程中信号系统之间结构和功能的高度保守性。