The glioma-associated protein SETA interacts with AIP1/Alix and AZIG-2 and modulates apoptosis in astrocytes
The glioma-associated protein SETA interacts with AIP1/Alix and AZIG-2 and modulates apoptosis in astrocytes
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DOI：
10.1074/jbc.m908994199
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发表时间：
2000-06-23
影响因子：
4.8
通讯作者：
Bogler, O
中科院分区：
文献类型：
--
作者：
Chen, B;Borinstein, SC;Bogler, O
在致瘤性星形胶质细胞(SETA)中表达的ARE同源3(SH3)结构域的表达与星形胶质细胞的致瘤状态有关。SETA编码多种含有一个或两个SH3结构域的适配蛋白，根据分离的cDNA序列的异质性，利用酵母双杂交筛选的神经胶质前体细胞cDNA文库中的SH3结构域，我们分离了大鼠ALG-2相互作用蛋白1或ALG-2相互作用蛋白X(AIP1/Alix)的同源物，体外对抗实验表明，SETA的SH3-N结构域与AIP1的富含Pro的C末端相互作用，在免疫共沉淀实验中，SETA和AIP1相互作用并可与凋亡相关基因2蛋白形成复合体，内源性SETA和AIP1蛋白在原代培养的大鼠星形胶质细胞中呈现相似的染色模式，多种SETA蛋白亚型在这些星形胶质细胞中的错误表达表明它们定位于肌动蛋白细胞骨架。此外，含有SH3-N结构域的SETA蛋白能够增强星形胶质细胞对紫外线诱导的细胞凋亡的敏感性。在这些实验中，分离的SH3-N结构域的表达影响最大，表明干扰内源性SETA和AIP1之间的相互作用使星形胶质细胞对DNA损伤的凋亡敏感。
Expression of the are 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.