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在酿酒酵母中,5.8S rRNA 加工的最后一步发生在细胞质中。

The final step in 5.8S rRNA processing is cytoplasmic in Saccharomyces cerevisiae.

机构信息

Wellcome Trust Centre for Cell Biology, University of Edinburgh, Edinburgh EH9 3JR, Scotland, UK.

出版信息

Mol Cell Biol. 2010 Feb;30(4):976-84. doi: 10.1128/MCB.01359-09. Epub 2009 Dec 14.

Abstract

The 18S rRNA component of yeast (Saccharomyces cerevisiae) 40S ribosomes undergoes cytoplasmic 3' cleavage following nuclear export, whereas exported pre-60S subunits were believed to contain only mature 5.8S and 25S rRNAs. However, in situ hybridization detected 3'-extended forms of 5.8S rRNA in the cytoplasm, which were lost when Crm1-dependent preribosome export was blocked by treatment with leptomycin B (LMB). LMB treatment rapidly blocked processing of 6S pre-rRNA to 5.8S rRNA, leading to TRAMP-dependent pre-rRNA degradation. The 6S pre-rRNA was coprecipitated with the 60S export adapter Nmd3 and cytoplasmic 60S synthesis factor Lsg1. The longer 5.8S+30 pre-rRNA (a form of 5.8S rRNA 3' extended by approximately 30 nucleotides) is processed to 6S by the nuclear exonuclease Rrp6, and nuclear pre-rRNA accumulated in the absence of Rrp6. In contrast, 6S to 5.8S processing requires the cytoplasmic exonuclease Ngl2, and cytoplasmic pre-rRNA accumulated in strains lacking Ngl2. We conclude that nuclear pre-60S particles containing the 6S pre-rRNA bind Nmd3 and Crm1 and are exported to the cytoplasm prior to final maturation by Ngl2.

摘要

酵母(酿酒酵母)40S 核糖体的 18S rRNA 成分在核输出后经历细胞质 3' 切割,而出口的前 60S 亚基被认为仅含有成熟的 5.8S 和 25S rRNA。然而,原位杂交在细胞质中检测到 5.8S rRNA 的 3' 延伸形式,当用莱普霉素 B(LMB)阻断 Crm1 依赖性前核糖体出口时,这些形式会丢失。LMB 处理迅速阻断了 6S 前 rRNA 向 5.8S rRNA 的加工,导致 TRAMP 依赖性前 rRNA 降解。6S 前 rRNA 与 60S 出口适配器 Nmd3 和细胞质 60S 合成因子 Lsg1 共沉淀。较长的 5.8S+30 前 rRNA(5.8S rRNA 的 3' 延伸约 30 个核苷酸的一种形式)被核外切酶 Rrp6 加工成 6S,并且在没有 Rrp6 的情况下核前 rRNA 积累。相比之下,6S 到 5.8S 的加工需要细胞质外切酶 Ngl2,并且在缺乏 Ngl2 的菌株中细胞质前 rRNA 积累。我们得出结论,含有 6S 前 rRNA 的核前 60S 颗粒与 Nmd3 和 Crm1 结合,并在 Ngl2 进行最终成熟之前被出口到细胞质。

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