Stoss O, Olbrich M, Hartmann A M, Konig H, Memmott J, Andreadis A, Stamm S
Max-Planck Institute of Neurobiology, Am Klopferspitz 18a, D-82152 Martinsried, the Forschungszentrum Karlsruhe, Institut für Genetik, Postfach 3640, D-76021 Karlsruhe, Germany.
J Biol Chem. 2001 Mar 23;276(12):8665-73. doi: 10.1074/jbc.M006851200. Epub 2000 Dec 15.
We identified the rat Sam68-like mammalian protein (rSLM-2), a member of the STAR (signal transduction and activation of RNA) protein family as a novel splicing regulatory protein. Using the yeast two-hybrid system, coimmunoprecipitations, and pull-down assays, we demonstrate that rSLM-2 interacts with various proteins involved in the regulation of alternative splicing, among them the serine/arginine-rich protein SRp30c, the splicing-associated factor YT521-B and the scaffold attachment factor B. rSLM-2 can influence the splicing pattern of the CD44v5, human transformer-2beta and tau minigenes in cotransfection experiments. This effect can be reversed by rSLM-2-interacting proteins. Employing rSLM-2 deletion variants, gel mobility shift assays, and linker scan mutations of the CD44 minigene, we show that the rSLM-2-dependent inclusion of exon v5 of the CD44 pre-mRNA is dependent on a short purine-rich sequence. Because the related protein of rSLM-2, Sam68, is believed to play a role as an adapter protein during signal transduction, we postulate that rSLM-2 is a link between signal transduction pathways and pre-mRNA processing.
我们鉴定出大鼠Sam68样哺乳动物蛋白（rSLM - 2），它是STAR（信号转导与RNA激活）蛋白家族的成员，是一种新型的剪接调节蛋白。利用酵母双杂交系统、免疫共沉淀和下拉实验，我们证明rSLM - 2与多种参与可变剪接调控的蛋白相互作用，其中包括富含丝氨酸/精氨酸的蛋白SRp30c、剪接相关因子YT521 - B和支架附着因子B。在共转染实验中，rSLM - 2可以影响CD44v5、人transformer - 2β和tau小基因的剪接模式。这种效应可以被与rSLM - 2相互作用的蛋白逆转。通过使用rSLM - 2缺失变体、凝胶迁移率变动分析以及CD44小基因的接头扫描突变，我们表明rSLM - 2依赖的CD44前体mRNA外显子v5的包含取决于一个短的富含嘌呤的序列。由于rSLM - 2的相关蛋白Sam68被认为在信号转导过程中作为衔接蛋白发挥作用，我们推测rSLM - 2是信号转导途径与前体mRNA加工之间的一个连接。
