Seykora John T, Mei Lijuan, Dotto G Paolo, Stein Paul L
Department of Dermatology, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA.
J Biol Chem. 2002 Jan 25;277(4):2812-22. doi: 10.1074/jbc.M106813200. Epub 2001 Nov 15.
The Src family tyrosine kinase, Fyn, can facilitate regulation of cell proliferation and differentiation. Mice with mutations in the fyn gene have defects in the brain, immune system, and epidermal differentiation. To identify molecules that may interact with Fyn in the epidermis, we performed a yeast two-hybrid interaction screen of a murine keratinocyte library. A novel adaptor-like molecule was isolated and termed Srcasm for Src activating and signaling molecule. Murine Srcasm is a 52.7-kDa protein that contains a VHS membrane association domain and a number of tyrosine motifs suggesting that it may be a substrate for Src family kinases and serve as an adaptor protein. Northern blot analysis of murine tissues demonstrates that Srcasm expression is highest in brain and kidney. In situ hybridization analysis reveals that srcasm mRNA is expressed in regions of the epidermis and hair follicle where keratinocyte differentiation occurs. In the brain, srcasm mRNA distribution correlates with that of fyn, with both being highly expressed in the hippocampal and cerebellar Purkinje neurons. Fyn can phosphorylate Srcasm, and association of these molecules relies on cooperative binding between the SH2 and SH3 domains of Fyn and corresponding canonical binding sites in Srcasm. Srcasm is capable of interacting with Grb2 and the regulatory subunit of phosphoinositide 3-kinase, p85, in a phosphorylation-dependent manner. The evidence suggests that Srcasm may help promote Src family kinase signaling in cells.
Src家族酪氨酸激酶Fyn可促进细胞增殖和分化的调控。fyn基因发生突变的小鼠在大脑、免疫系统和表皮分化方面存在缺陷。为了鉴定可能在表皮中与Fyn相互作用的分子,我们对小鼠角质形成细胞文库进行了酵母双杂交相互作用筛选。分离出一种新型的类似接头分子,命名为Srcasm,即Src激活和信号分子。小鼠Srcasm是一种52.7 kDa的蛋白质,含有一个VHS膜结合结构域和多个酪氨酸基序,这表明它可能是Src家族激酶的底物,并作为一种接头蛋白发挥作用。对小鼠组织的Northern印迹分析表明,Srcasm在大脑和肾脏中的表达最高。原位杂交分析显示,srcasm mRNA在角质形成细胞分化发生的表皮和毛囊区域表达。在大脑中,srcasm mRNA的分布与fyn的分布相关,二者在海马和小脑浦肯野神经元中均高表达。Fyn可使Srcasm磷酸化,这些分子的结合依赖于Fyn的SH2和SH3结构域与Srcasm中相应的典型结合位点之间的协同结合。Srcasm能够以磷酸化依赖的方式与Grb2和磷酸肌醇3激酶的调节亚基p85相互作用。证据表明,Srcasm可能有助于促进细胞中Src家族激酶的信号传导。