Critical Role for Saccharomyces cerevisiae Asc1p in Translational Initiation at Elevated Temperatures.
Critical Role for Saccharomyces cerevisiae Asc1p in Translational Initiation at Elevated Temperatures.
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DOI：
10.1002/pmic.201800208
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发表时间：
2018-12
期刊：
影响因子：
3.4
通讯作者：
Link AJ
中科院分区：
文献类型：
--
作者：
Gerbasi VR;Browne CM;Samir P;Shen B;Sun M;Hazelbaker DZ;Galassie AC;Frank J;Link AJ
真核核糖体蛋白RACK 1/Asc 1 p定位于40 S亚基的mRNA出口通道，但在mRNA翻译中缺乏确定的作用。ASC 1缺陷的酿酒酵母表现出温度敏感性生长。使用这个无效突变体，我们评估了Asc 1 p在翻译和核糖体生物合成中的潜在作用。我们发现在限制性温度下asc 1 Δ无效突变体的多聚核糖体减少。为了测试Asc 1 p在核糖体稳定性中的作用，我们使用冷冻电子显微镜在允许和非允许温度下检查asc 1 Δ酵母缺失突变体中80 s核糖体的结构。cryoEM表明Asc 1 p的丢失不会严重破坏这种复杂结构的形成。我们在asc 1 Δ无效突变体中没有发现rRNA加工缺陷。我们应用蛋白质组学方法来调查Asc 1 p损失对酵母蛋白质全局翻译的影响。在非允许温度下，asc 1 Δ突变体的核糖体蛋白和其他对翻译至关重要的因子水平降低。总的来说，这些结果与最近的观察结果一致，表明Asc 1 p对短mRNA的核糖体占据很重要。我们的研究结果表明，Asc 1核糖体蛋白在热应激期间的翻译是至关重要的。
The eukaryotic ribosomal protein RACK1/Asc1p is localized at the mRNA exit channel of the 40S subunit but lacks a defined role in mRNA translation. Saccharomyces cerevisiae deficient in ASC1 exhibit temperature-sensitive growth. Using this null mutant, we evaluated potential roles for Asc1p in translation and ribosome biogenesis. We found that at the restrictive temperature the asc1Δ null mutant had reduced polyribosomes. To test the role of Asc1p in ribosome stability, we used cryoelectron microscopy to examine the structure of 80s ribosomes in an asc1Δ yeast deletion mutant at both the permissive and nonpermissive temperatures. cryoEM indicated that loss of Asc1p does not severely disrupt formation of this complex structure. We found no defect in rRNA processing in the asc1Δ null mutant. We applied a proteomic approach to survey the effect of Asc1p loss on the global translation of yeast proteins. At the nonpermissive temperature, the asc1Δ mutant had reduced levels of ribosomal proteins and other factors critical for translation. Collectively, these results are consistent with recent observations suggesting that Asc1p is important for ribosome occupancy of short mRNAs. Our results show the Asc1 ribosomal protein is critical in translation during heat stress.
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影响因子：
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作者：
Langlois, Robert;Pallesen, Jesper;Ash, Jordan T.;Ho, Danny Nam;Rubinstein, John L.;Frank, Joachim
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