BimL directly neutralizes Bcl-xL to promote Bax activation during UV-induced apoptosis
BimL directly neutralizes Bcl-xL to promote Bax activation during UV-induced apoptosis
复制标题
BimL 直接中和 Bcl-xL，促进 UV 诱导细胞凋亡过程中的 Bax 激活
DOI：
10.1016/j.febslet.2009.04.045
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发表时间：
2009-06-18
期刊：
影响因子：
3.5
通讯作者：
Chen, Wei R.
中科院分区：
文献类型：
--
作者：
Wang, Xianwang;Xing, Da;Chen, Wei R.
Bcl-2-interacting mediator of cell death（Bim）被认为是通过Bax激活启动内源性凋亡途径。以前的研究表明，BimL参与了紫外线诱导的细胞凋亡，但目前尚不清楚Bim是否通过直接参与或释放它从促生存的亲戚，如Bcl-xL激活Bax。在本研究中，我们试图确定BimL和Bax/Bcl-xL之间的相互作用在紫外线（UV）诱导的细胞凋亡。在我们的实验中，BimL激活似乎是一个重要的事件，如通过RNAi方法敲低内源性BimL显著抑制细胞死亡、caspase-3活性和Bax易位所证明的。荧光共振能量转移（FRET）和免疫共沉淀（CO-IP）分析表明Bcl-xL直接与Bax结合以抑制其活化，而BimL直接中和Bcl-xL，然后在UV照射下释放Bax并活化。在我们的实验中没有检测到BimL和Bax之间的相互作用，无论是在活细胞中使用FRET方法还是内源性CO-IP测定。因此，我们的研究结果首次在活细胞中提供了强有力的证据，证明BimL在紫外线照射下通过废除Bcl-xL并促进Bax激活来启动细胞凋亡。结构摘要：MINT-7034091：BIML（uniprotkb：O 43521）物理相互作用（MI：0218）与Bcl 2-Xl（uniprotkb：Q92934）通过抗诱饵免疫共沉淀法（MI：0006）MINT-7034079：Bcl2-Xl（uniprotkb：Q92934）物理相互作用（MI：0218）与BAX（uniprotkb：Q 07812）和BIML（uniprotkb：O 43521）通过抗诱饵免疫共沉淀（MI：0006）MINT-7034069：BAX（uniprotkb：Q 07812）物理相互作用（MI：0218）与BIML（uniprotkb：O 43521）通过抗诱饵免疫共沉淀（MI：0006）MINT-7034114：BIML（uniprotkb：O 43521）和BAX（uniprotkb：Q 07812）通过荧光共振能量转移（MI：0055）发生物理相互作用（MI：0218）（C）2009欧洲生物化学学会联合会。Elsevier B. V.出版，保留所有权利。
Bcl-2-interacting mediator of cell death (Bim) has been considered to initiate intrinsic apoptotic pathway through Bax activation. Previous studies indicated that BimL was involved in UV-induced apoptosis, but it remains unclear whether Bim activates Bax by directly engaging it or by releasing it from pro-survival relatives such as Bcl-xL. In this study, we attempt to determine the interactions between BimL and Bax/Bcl-xL during Ultraviolet (UV)-induced apoptosis. BimL activation appeared to be an important event in our experiments, as demonstrated by the significant inhibition of cell death, caspase-3 activity, and Bax translocation in cells with knockdown of endogenous BimL by RNAi approach. Both fluorescence resonance energy transfer (FRET) and Co-immunoprecipitation (CO-IP) assays indicated that Bcl-xL directly bound to Bax to inhibit its activation, while BimL directly neutralized Bcl-xL, followed by Bax release and activation upon UV irradiation. Not detected in our experiment was the interaction between BimL and Bax either using FRET approach in living cells or endogenous CO-IP assay. Thus, our findings provide strong evidence in living cells for the first time that BimL initiates apoptosis by abrogating Bcl-xL and promoting Bax activation under UV irradiation.Structured summary:MINT-7034091: BIML (uniprotkb: O43521) physically interacts (MI: 0218) with Bcl2-Xl (uniprotkb: Q92934) by anti bait coimmunoprecipitation (MI: 0006)MINT-7034079: Bcl2-Xl (uniprotkb: Q92934) physically interacts (MI: 0218) with BAX (uniprotkb: Q07812) and BIML (uniprotkb: O43521) by anti bait coimmunoprecipitation (MI: 0006)MINT-7034069: BAX (uniprotkb: Q07812) physically interacts (MI: 0218) with BIML (uniprotkb: O43521) by anti bait coimmunoprecipitation (MI: 0006)MINT-7034114: BIML (uniprotkb: O43521) and BAX (uniprotkb: Q07812) physically interact (MI: 0218) by fluorescent resonance energy transfer (MI: 0055) (C) 2009 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.