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PMID: 10411134 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Yeast Rnt1p is required for cleavage of the pre-ribosomal RNA in the 3' ETS but not the 5' ETS.

RNA (New York, N.Y.) ·Vol. 5 ·No. 7 ·1999-07-00 ·Pages 909-17

Kufel J, Dichtl B, Tollervey D

Abstract

We have reexamined the role of yeast RNase III (Rnt1p) in ribosome synthesis. Analysis of pre-rRNA processing in a strain carrying a complete deletion of the RNT1 gene demonstrated that the absence of Rnt1p does not block cleavage at site A0 in the 5' external transcribed spacers (ETS), although the early pre-rRNA cleavages at sites A0, A1, and A2 are kinetically delayed. In contrast, cleavage in the 3' ETS is completely inhibited in the absence of Rnt1p, leading to the synthesis of a reduced level of a 3' extended form of the 25S rRNA. The 3' extended forms of the pre-rRNAs are consistent with the major termination at site T2 (+210). We conclude that Rnt1p is required for cleavage in the 3' ETS but not for cleavage at site A0. The sites of in vivo cleavage in the 3' ETS were mapped by primer extension. Two sites of Rnt1p-dependent cleavage were identified that lie on opposite sides of a predicted stem loop structure, at +14 and +49. These are in good agreement with the consensus Rnt1p cleavage site. Processing of the 3' end of the mature 25S rRNA sequence in wild-type cells was found to occur concomitantly with processing of the 5' end of the 5.8S rRNA, supporting previous proposals that processing in ITS1 and the 3' ETS is coupled.

MeSH Terms
Base Sequence Endoribonucleases/metabolism Fungal Proteins/metabolism Hydrolysis Molecular Sequence Data Nucleic Acid Conformation RNA Precursors/chemistry,metabolism RNA Processing, Post-Transcriptional RNA, Ribosomal/chemistry,metabolism Ribonuclease III
Chemicals
Fungal Proteins RNA Precursors RNA, Ribosomal Endoribonucleases Ribonuclease III
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Kufel J
Institute of Cell and Molecular Biology, University of Edinburgh, United Kingdom.
Dichtl B
Tollervey D
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Article Info
Journal
RNA (New York, N.Y.)
Abbr.
RNA
ISSN
1355-8382
DOI
10.1017/s135583829999026x
Published
1999-07-00
Pages
909-17
Language
English
Region
United States
NLM ID
9509184
PMCID
PMC1369815
Subset
IM
Grants
Wellcome Trust · United Kingdom
External Links
PubMed source DOI: 10.1017/s135583829999026x PMC Full Text View journal details
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