Ubiquitination of stalled ribosomes enables mRNA decay via HBS-1 and NONU-1 in vivo.
Ubiquitination of stalled ribosomes enables mRNA decay via HBS-1 and NONU-1 in vivo.
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DOI：
10.1371/journal.pgen.1010577
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发表时间：
2023-01
期刊：
影响因子：
4.5
通讯作者：
中科院分区：
文献类型：
--
作者：
当核糖体翻译遗传密码时，它们可能会遇到各种阻碍其进展的障碍。如果核糖体停滞很长时间，细胞会由于翻译核糖体的损失和异常蛋白质产物的积累而受损。因此，为了保护细胞，停滞的核糖体经历了一系列反应来缓解停滞并降解冒犯的mRNA，这一过程称为No-Go mRNA Decay（NGD）。虽然NGD的机制很多都是已知的，但沿着这条途径的事件和因素的精确顺序还没有得到测试。在这里，我们部署C。elegans来解开包括NGD的协调事件。利用一种新的报告和正向和反向遗传学，我们确定了NGD所需的机制。我们随后的分子分析定义了至少两个核糖体蛋白（eS 10和uS 10）的泛素化的功能要求，我们发现，eS 10和uS 10上缺乏泛素化位点的核糖体不能在体内进行NGD。我们表明，核酸酶NONU-1的行为后，泛素连接酶ZNF-598，并发现一个新的要求核糖体救援因子HBS-1/PELO-1的mRNA衰变通过NONU-1。总之，我们的工作表明核糖体信号的mRNA抑制效应器的机制，我们描绘了一个明确的NGD通路工作的抑制因子之间的联系。核糖体是大型分子机器，被赋予阅读mRNA和构建蛋白质的基本任务。核糖体不受阻碍地继续工作是至关重要的，以避免mRNA上核糖体之间的交通堵塞;然而，它们有时会遇到挑战，使交通堵塞不可避免。在这些情况下，细胞已经进化出了清理停滞的核糖体和去除有害mRNA的机制。这种机制的大部分现在已经确定，但因素之间的关系还有待检验。在这里，在C。我们识别出对失速做出反应的机制，并对因子进行排序。我们发现，核糖体必须标记的泛素信号mRNA衰变，因为核糖体缺乏泛素网站不能引起mRNA衰变。此外，我们发现，这些泛素信号招募酶，削减mRNA，我们提出的证据表明，核糖体救援因子所需的mRNA衰变反应以及。总的来说，我们的工作提供了协调消除阻碍核糖体的有问题的mRNA的因素之间的新联系。
As ribosomes translate the genetic code, they can encounter a variety of obstacles that hinder their progress. If ribosomes stall for prolonged times, cells suffer due to the loss of translating ribosomes and the accumulation of aberrant protein products. Thus to protect cells, stalled ribosomes experience a series of reactions to relieve the stall and degrade the offending mRNA, a process known as No-Go mRNA Decay (NGD). While much of the machinery for NGD is known, the precise ordering of events and factors along this pathway has not been tested. Here, we deploy C. elegans to unravel the coordinated events comprising NGD. Utilizing a novel reporter and forward and reverse genetics, we identify the machinery required for NGD. Our subsequent molecular analyses define a functional requirement for ubiquitination on at least two ribosomal proteins (eS10 and uS10), and we show that ribosomes lacking ubiquitination sites on eS10 and uS10 fail to perform NGD in vivo. We show that the nuclease NONU-1 acts after the ubiquitin ligase ZNF-598, and discover a novel requirement for the ribosome rescue factors HBS-1/PELO-1 in mRNA decay via NONU-1. Taken together, our work demonstrates mechanisms by which ribosomes signal to effectors of mRNA repression, and we delineate links between repressive factors working toward a well-defined NGD pathway. Ribosomes are large molecular machines entrusted with the essential task of reading mRNAs and building proteins. It is critical that ribosomes proceed with their work undeterred, so as to avoid traffic jams between ribosomes on mRNAs; however, they sometimes experience challenges which make traffic jams unavoidable. In these cases, cells have evolved machinery to clean up stalled ribosomes and remove offending mRNAs. Much of this machinery is now identified, but the relationships between factors has yet to be tested. Here, in C. elegans we identify the machinery that responds to stalls, and we order factors relative to one another. We find that ribosomes must be tagged by ubiquitin to signal mRNA decay, as ribosomes lacking ubiquitin sites fail to elicit mRNA decay. Furthermore, we find that these ubiquitin signals recruit an enzyme which cuts mRNA, and we present evidence that ribosome rescue factors are required for the mRNA decay reaction as well. Overall, our work provides new connections between factors orchestrating the elimination of problematic mRNAs that stall ribosomes.
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影响因子：
10.5
作者：
BEJSOVEC, A;ANDERSON, P
通讯作者：
ANDERSON, P
影响因子：
64.8
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Hassabis D
影响因子：
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Dey G;Meyer T
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影响因子：
8.8
作者：
Glover, Marissa L.;Burroughs, A. Max;Arribere, Joshua A.
通讯作者：
Arribere, Joshua A.
影响因子：
11.4
作者：
Alam, SL;Sun, J;Sundquist, WI
通讯作者：
Sundquist, WI