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酵母前体rRNA 5' 外部转录间隔区中U3 snoRNA关键结合位点的突变分析。

Mutational analysis of an essential binding site for the U3 snoRNA in the 5' external transcribed spacer of yeast pre-rRNA.

作者信息

Beltrame M, Henry Y, Tollervey D

机构信息

Dipartimento di Genetica e di Biologia dei Microrganismi, Università degli Studi di Milano, Italy.

出版信息

Nucleic Acids Res. 1994 Oct 11;22(20):4057-65. doi: 10.1093/nar/22.20.4057.

DOI:10.1093/nar/22.20.4057
PMID:7937130
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC331890/
Abstract

The small nucleolar RNA U3 is essential for viability in yeast. We have previously shown that U3 can be cross-linked in vivo to the pre-rRNA in the 5' external transcribed spacer (ETS), at +470. This ETS region contains 10 nucleotides of perfect complementarity to U3. In a genetic background where the mutated rDNA is the only transcribed rDNA repeat, the deletion of the 10 nt complementary to U3 is lethal. Cells lacking the U3 complementary sequence in pre-rRNA fail to accumulate 18S rRNA: pre-rRNA processing is inhibited at sites A0 in the 5' ETS, A1 at the 5' end of 18S rRNA and A2 in ITS1. We show here that effects on processing at site A0 are specific for U3 and its associated proteins and are not seen on depletion of other snoRNP components. The deletion of the sequence complementary to U3 in the ETS therefore mimics all the known effects of the depletion of U3 in trans. This indicates that we have identified an essential U3 binding site on pre-rRNA, required in cis for the maturation of 18S rRNA.

摘要

小核仁RNA U3对酵母的生存能力至关重要。我们之前已经表明，U3在体内可与5'外部转录间隔区（ETS）中位于+470处的前体rRNA发生交联。该ETS区域包含与U3完全互补的10个核苷酸。在突变的rDNA是唯一转录的rDNA重复序列的遗传背景下，删除与U3互补的10个核苷酸是致死的。前体rRNA中缺乏U3互补序列的细胞无法积累18S rRNA：前体rRNA加工在5' ETS的A0位点、18S rRNA 5'端的A1位点和ITS1中的A2位点受到抑制。我们在此表明，对A0位点加工的影响是U3及其相关蛋白所特有的，在其他snoRNP成分缺失时未见此现象。因此，ETS中与U3互补序列的缺失模拟了反式缺失U3的所有已知效应。这表明我们已经在pre-rRNA上鉴定出一个必需的U3结合位点，它是18S rRNA顺式成熟所必需的。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/081c/331890/d9ee4e638ef1/nar00044-0054-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/081c/331890/c6fe18caf3e3/nar00044-0052-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/081c/331890/f455ad94b52e/nar00044-0052-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/081c/331890/8d8ae9072776/nar00044-0052-c.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/081c/331890/72379b83c68d/nar00044-0053-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/081c/331890/367e5fd32350/nar00044-0054-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/081c/331890/d9ee4e638ef1/nar00044-0054-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/081c/331890/c6fe18caf3e3/nar00044-0052-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/081c/331890/f455ad94b52e/nar00044-0052-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/081c/331890/8d8ae9072776/nar00044-0052-c.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/081c/331890/72379b83c68d/nar00044-0053-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/081c/331890/367e5fd32350/nar00044-0054-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/081c/331890/d9ee4e638ef1/nar00044-0054-b.jpg

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