RhoB induces apoptosis via direct interaction with TNFAIP1 in HeLa cells
RhoB induces apoptosis via direct interaction with TNFAIP1 in HeLa cells
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DOI：
10.1002/ijc.24617
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发表时间：
2009-12-01
影响因子：
6.4
通讯作者：
Seo, Yeon-Soo
中科院分区：
文献类型：
--
作者：
Kim, Dong-Myung;Chung, Kyung-Sook;Seo, Yeon-Soo
RhoB是一种肿瘤抑制因子，已经成为一种有趣的癌症靶点，并且已经进行了旨在了解其在细胞凋亡中的作用的广泛研究。在我们的研究中，我们调查了RhoB相互作用分子在细胞凋亡中的参与。为了鉴定RhoB相互作用蛋白，我们使用RhoB作为诱饵进行酵母双杂交筛选试验，并分离出TNFAIP 1，一种含有BTB/POZ结构域的TNF α诱导蛋白。RhoB和TNFAIP 1之间的相互作用在体内通过免疫共沉淀研究和体外结合试验证明。RFP-TNFAIP 1与EGFP-RhoB部分共定位。RIII和TNFAIP 1在内体中的部分共定位表明RhoB-TNFAIP 1相互作用可能在细胞凋亡中具有功能性作用。TNFAIP 1引起促凋亡活性，而RhoB和TNFAIP 1的同时表达导致HeLa细胞凋亡的急剧增加。此外，使用siRNA敲低RhoB清楚地从TNFAIP 1诱导的细胞凋亡中拯救细胞。这一发现表明RhoB和TNFAIP 1之间的相互作用对于诱导HeLa细胞凋亡至关重要。在凋亡细胞中增加的SAPK/JNK磷酸化的观察和JNK抑制剂抑制凋亡的发现表明SAPK/JNK信号传导可能参与RhoB-TNFAIP 1相互作用诱导的凋亡。总之，我们发现RhoB与TNFAIP 1相互作用，通过SAPK/JNK介导的信号转导机制调节细胞凋亡。(c)2009年UICC
RhoB, a tumor suppressor, has emerged as an interesting cancer target, and extensive studies aimed at understanding its role in apoptosis have been performed. In our study, we investigated the involvement of RhoB-interacting molecules in apoptosis. To identify RhoB-interacting proteins, we performed yeast-two hybrid screening assays using RhoB as a bait and isolated TNFAIP1, a TNF alpha-induced protein containing the BTB/POZ domain. The interaction between RhoB and TNFAIP1 was demonstrated in vivo through coimmunoprecipitation studies and in vitro binding assays. RFP-TNFAIP1 was found to be partially colocalized with EGFP-RhoB. The partial colocalization of Rill and TNFAIP1 in endosomes suggests that RhoB-TNFAIP1 interactions may have a functional role in apoptosis. TNFAIP1 elicited proapoptotic activity, while simultaneous expression of RhoB and TNFAIP1 resulted in a dramatic increase in apoptosis in HeLa cells. Furthermore, knockdown of RhoB using siRNA clearly rescued cells from apoptosis induced by TNFAIP1. This finding suggests that interactions between RhoB and TNFAIP1 are crucial for induction of apoptosis in HeLa cells. The observation of increased SAPK/JNK phosphorylation in apoptotic cells and the finding that a JNK inhibitor suppressed apoptosis indicates that SAPK/JNK signaling may be involved in apoptosis induced by RhoB-TNFAIP1 interactions. In conclusion, we found that RhoB interacts with TNFAIP1 to regulate apoptosis via a SAPK/JNK-mediated signal transduction mechanism. (c) 2009 UICC