Regulation of c-Fes tyrosine kinase activity by coiled-coil and SH2 domains: analysis with Saccharomyces cerevisiae.
Regulation of c-Fes tyrosine kinase activity by coiled-coil and SH2 domains: analysis with Saccharomyces cerevisiae.
复制标题
卷曲螺旋和 SH2 结构域对 c-Fes 酪氨酸激酶活性的调节：酿酒酵母分析。
DOI：
10.1021/bi0272499
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发表时间：
2003
期刊：
影响因子：
2.9
通讯作者：
Smithgall,ThomasE
中科院分区：
文献类型：
--
作者：
Takashima,Yoshio;Delfino,FrankJ;Engen,JohnR;Superti-Furga,Giulio;Smithgall,ThomasE
c-Fes蛋白酪氨酸激酶调节多种细胞类型的生长和分化，包括骨髓造血细胞、血管内皮细胞和神经元。在结构上，Fes是由一个独特的N-末端区域与卷曲螺旋寡聚化基序，其次是SH 2和激酶结构域。虽然Fes激酶活性在细胞中受到严格调控，但其负调控的结构基础尚不清楚。在这篇报道中，c-Fes在酵母中表达，以确定调节是激酶内在的还是依赖于哺乳动物细胞中发现的蛋白质因子。野生型Fes激酶活性在酵母中被完全抑制，并且不影响细胞生长。突变或删除更多的N-末端c-Fes卷曲螺旋结构域逆转负调控，导致强烈的激酶激活和抑制酵母细胞生长。类似地，用v-Src替换野生型SH 2结构域诱导强激酶活化和生长抑制表型。磷酸特异性抗体的免疫印迹显示，通过任一机制激活Fes诱导激活环酪氨酸残基（Tyr 713）的自磷酸化。这些数据支持的想法，Fes在体内自然地采取非活性构象，和非活性结构的维持需要卷曲螺旋和SH 2结构域。
The c-Fes protein-tyrosine kinase regulates the growth and differentiation of diverse cell types, including myeloid hematopoietic cells, vascular endothelial cells, and neurons. Structurally, Fes is composed of a unique N-terminal region with coiled-coil oligomerization motifs, followed by SH2 and kinase domains. Although Fes kinase activity is tightly regulated in cells, the structural basis for its negative regulation is not clear. In this report, c-Fes was expressed inSaccharomyces cerevisiaeto determine whether regulation is kinase-intrinsic or dependent upon protein factors found in mammalian cells. Wild-type Fes kinase activity was completely repressed in yeast and did not affect cell growth. Mutation or deletion of the more N-terminal c-Fes coiled-coil domain reversed negative regulation, leading to strong kinase activation and suppression of yeast cell growth. Similarly, replacement of the wild-type SH2 domain with that of v-Src induced strong kinase activation and the growth-inhibitory phenotype. Immunoblotting with phosphospecific antibodies shows that activation of Fes by either mechanism induced autophosphorylation of the activation loop tyrosine residue (Tyr 713). These data support the idea that Fes naturally adopts an inactive conformation in vivo, and that maintenance of the inactive structure requires the coiled-coil and SH2 domains.