Peptides derived from the bifunctional kinase/RNase enzyme IRE1α modulate IRE1α activity and protect cells from endoplasmic reticulum stress
Peptides derived from the bifunctional kinase/RNase enzyme IRE1α modulate IRE1α activity and protect cells from endoplasmic reticulum stress
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DOI：
10.1096/fj.11-182931
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发表时间：
2011-09-01
期刊：
影响因子：
4.8
通讯作者：
Chevet, Eric
中科院分区：
文献类型：
--
作者：
Bouchecareilh, Marion;Higa, Arisa;Chevet, Eric
双功能激酶/RNA酶IRE 1 α的激活是内质网（ER）中错误折叠蛋白积累引发的适应性反应的一部分。为了促进ER稳态的恢复，IRE 1 α分子寡聚化，允许其转自磷酸化和核糖核酸内切酶活化。这些反过来又诱导特定转录和转录后程序的激活。为了鉴定IRE 1 α活性的新型和选择性调节剂，我们使用通过基于活性的体外测定鉴定的IRE 1 α衍生肽研究IRE 1 α寡聚化特性。然后，我们使用这些肽在体外和体内使用培养的人肝细胞癌衍生的HuH 7细胞和秀丽隐杆线虫实验系统来探测IRE 1 α活性。我们鉴定了来自人IRE 1 α激酶结构域的肽，其在体外促进IRE 1 α寡聚化，在体外（1.7 x）在细胞培养物（1.8 x）和体内（1.3 x）增强其Xbp 1 mRNA切割活性，并减弱ER应激介导的JNK激活和调节IRE 1依赖性mRNA衰减（RIDD）。这伴随着衣霉素诱导的ER应激的存活率增加2.5倍，并且在表达该肽的细胞中凋亡减少1.4倍。因此，靶向和选择性激活IRE 1 α的催化特性可能因此定义新的策略，以保护细胞免受ER应激信号传导的有害影响。Bouchecareilh，M.，Higa，A.，弗里堡，南，Moenner，M.，Chevet，E.衍生自双功能激酶/RNA酶IRE 1 α的肽调节IRE 1 α活性并保护细胞免受内质网应激。FASEB J. 25，3115-3129（2011）。www.fasebj.org
Activation of the bifunctional kinase/RNase enzyme IRE1 alpha is part of an adaptive response triggered on accumulation of misfolded proteins in the endoplasmic reticulum (ER). To facilitate recovery of ER homeostasis, IRE1 alpha molecules oligomerize, allowing for their transautophosphorylation and endoribonuclease activation. These, in turn, induce the activation of specific transcriptional and post-transcriptional programs. To identify novel and selective modulators of IRE1 alpha activity, we investigated IRE1 alpha oligomerization properties using IRE1 alpha-derived peptides identified through an activity-based in vitro assay. We then used these peptides to probe IRE1 alpha activity in vitro and in vivo using both cultured human hepatocellular carcinoma-derived HuH7 cells and Caenorhabditis elegans experimental systems. We identified a peptide derived from the kinase domain of human IRE1 alpha, which promoted IRE1 alpha oligomerization in vitro, enhanced its Xbp1 mRNA cleavage activity in vitro (1.7 x) in cell culture (1.8 x) and in vivo (1.3 x), and attenuated both ER stress-mediated JNK activation and regulated IRE1-dependent mRNA decay (RIDD). This was accompanied by a 2.5-fold increase in survival on tunicamycin-induced ER stress and reduced apoptosis by 1.4-fold in cells expressing this peptide. Hence, targeted and selective activation of the catalytic properties of IRE1 alpha may consequently define new strategies to protect cells from deleterious effects of ER stress signaling.-Bouchecareilh, M., Higa, A., Fribourg, S., Moenner, M., Chevet, E. Peptides derived from the bifunctional kinase/RNase enzyme IRE1 alpha modulate IRE1 alpha activity and protect cells from endoplasmic reticulum stress. FASEB J. 25, 3115-3129 (2011). www.fasebj.org