The Glioma-associated Protein SETA Interacts with AIP1/Alix and ALG-2 and Modulates Apoptosis in Astrocytes*
The Glioma-associated Protein SETA Interacts with AIP1/Alix and ALG-2 and Modulates Apoptosis in Astrocytes*
复制标题
胶质瘤相关蛋白 SETA 与 AIP1/Alix 和 ALG-2 相互作用并调节星形胶质细胞的细胞凋亡*
DOI：
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发表时间：
2000
影响因子：
4.8
通讯作者：
O. Bogler
中科院分区：
文献类型：
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作者：
Baihua Chen;S. Borinstein;J. Gillis;V. Sykes;O. Bogler
含src同源性3（SH 3）结构域的星形胶质细胞致瘤基因（SETA）的表达与星形胶质细胞的致瘤性有关。SETA编码含有一个或两个SH 3结构域的多种衔接蛋白，如分离的cDNA的序列异质性所示。在酵母双杂交筛选神经胶质祖细胞cDNA文库中使用两个SH 3结构域，我们分离出ALG-2相互作用蛋白1或ALG-2相互作用蛋白X（AIP 1/阿利克斯）的大鼠同源物。体外对抗实验表明，SETA的SH 3-N结构域与AIP 1富含脯氨酸的C端相互作用。在免疫共沉淀实验中，SETA和AIP 1相互作用，并能与糖尿病相关基因2蛋白形成复合物。内源性SETA和AIP 1蛋白在原代大鼠星形胶质细胞中显示出相似的染色模式。错误表达的各种SETA蛋白异构体在这些星形胶质细胞显示，它们定位于肌动蛋白细胞骨架。此外，含有SH 3-N结构域的SETA蛋白能够使星形胶质细胞对UV照射诱导的凋亡敏感。在这些实验中，分离的SH 3-N结构域的表达具有最大的影响，表明内源性SETA和AIP 1之间的相互作用的干扰使星形胶质细胞响应于DNA损伤而对凋亡敏感。
Expression of the src homology 3 (SH3) domain-containing expressed in tumorigenic astrocytes (SETA) gene is associated with the tumorigenic state in astrocytes. SETA encodes a variety of adapter proteins containing either one or two SH3 domains, as suggested by the sequence heterogeneity of isolated cDNAs. Using both SH3 domains in a yeast two-hybrid screen of a glial progenitor cell cDNA library, we isolated the rat homolog of the ALG-2-interacting protein 1 or ALG-2-interacting protein X (AIP1/Alix). In vitro confrontation experiments showed that the SH3-N domain of SETA interacted with the proline-rich C terminus of AIP1. In co-immunoprecipitation experiments, SETA and AIP1 interacted and could form a complex with apoptosis-linked gene 2 protein. Endogenous SETA and AIP1 proteins showed similar patterns of staining in primary rat astrocytes. Misexpression of a variety of SETA protein isoforms in these astrocytes revealed that they localized to the actin cytoskeleton. Furthermore, SETA proteins containing the SH3-N domain were able to sensitize astrocytes to apoptosis induced by UV irradiation. Expression of the isolated SH3-N domain had the greatest effect in these experiments, indicating that interference in the interaction between endogenous SETA and AIP1 sensitizes astrocytes to apoptosis in response to DNA damage.