Gcn2二聚体与大60S核糖体亚基复合物的结构

IF 9.1 1区 综合性期刊 Q1 MULTIDISCIPLINARY SCIENCES
Helge Paternoga, Lu Xia, Lyudmila Dimitrova-Paternoga, Sihan Li, Liewei L. Yan, Malte Oestereich, Sergo Kasvandik, Ankanahalli N. Nanjaraj Urs, Bertrand Beckert, Tanel Tenson, Hani Zaher, Toshifumi Inada, Daniel N. Wilson
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引用次数: 0

摘要

综合应激反应(integrated stress response, ISR)是真核细胞响应各种不同环境应激的中枢信号网络。这种应激引起核糖体碰撞,导致激酶Gcn2的激活,导致真核起始因子2的磷酸化和失活,从而促进mrna的选择性翻译以恢复体内平衡。尽管ISR的重要性和过去几十年的深入研究,Gcn2如何与核糖体颗粒相互作用的结构见解一直缺乏。利用离体亲和纯化方法,我们获得了酵母Gcn2二聚体与核糖体60S亚基复合物的低温电镜结构。Gcn2二聚体是由组氨酸tRNA合成酶样结构域二聚化形成的,它与60S亚基的茎基和肌毒素-蓖麻毒素环建立了广泛的相互作用。Gcn2的c端结构域也是二聚化的,占据60S亚基肽基转移酶中心的A和p位点tRNA结合位点。互补功能研究表明,Gcn2与60S亚基的结合不需要共激活剂Gcn1或Gcn20,也不会导致eIF2α的磷酸化。相反,在应激时,我们观察到Gcn2从60S亚基转移到碰撞核糖体片段中,这表明Gcn2 - 60S复合物代表一种非活性待机状态,能够快速重新分配到碰撞核糖体中,从而促进对应激的快速有效反应。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Structure of a Gcn2 dimer in complex with the large 60S ribosomal subunit
The integrated stress response (ISR) is a central signaling network that enables eukaryotic cells to respond to a variety of different environmental stresses. Such stresses cause ribosome collisions that lead to activation of the kinase Gcn2, resulting in the phosphorylation and inactivation of eukaryotic initiation factor 2 and thereby promoting selective translation of mRNAs to restore homeostasis. Despite the importance of the ISR and intensive study over the past decades, structural insight into how Gcn2 interacts with ribosomal particles has been lacking. Using ex vivo affinity purification approaches, we have obtained a cryoelectron microscopy structure of a yeast Gcn2 dimer in complex with the ribosomal 60S subunit. The Gcn2 dimer is formed by dimerization of the histidine tRNA synthetase-like domains, which establish extensive interactions with the stalk-base and sarcin–ricin loop of the 60S subunit. The C-terminal domain of Gcn2 is also dimerized and occupies the A- and P-site tRNA binding sites at the peptidyl-transferase center of the 60S subunit. Complementary functional studies indicate that binding of Gcn2 to the 60S subunit does not require the coactivators Gcn1 or Gcn20, nor does it lead to phosphorylation of eIF2α. Instead, upon stress, we observe a shift of Gcn2 from the 60S subunit into the colliding ribosome fraction, suggesting that the Gcn2–60S complex represents an inactive stand-by state to enable a rapid redistribution to collided ribosomes, and thereby facilitating a quick and efficient response to stress.
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来源期刊
CiteScore
19.00
自引率
0.90%
发文量
3575
审稿时长
2.5 months
期刊介绍: The Proceedings of the National Academy of Sciences (PNAS), a peer-reviewed journal of the National Academy of Sciences (NAS), serves as an authoritative source for high-impact, original research across the biological, physical, and social sciences. With a global scope, the journal welcomes submissions from researchers worldwide, making it an inclusive platform for advancing scientific knowledge.
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