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PMID: 25125595 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

A cluster of methylations in the domain IV of 25S rRNA is required for ribosome stability.

RNA (New York, N.Y.) ·Vol. 20 ·No. 10 ·2014-10-00 ·Pages 1632-44

Gigova A, Duggimpudi S, Pollex T, Schaefer M, Koš M

Abstract

In all three domains of life ribosomal RNAs are extensively modified at functionally important sites of the ribosome. These modifications are believed to fine-tune the ribosome structure for optimal translation. However, the precise mechanistic effect of modifications on ribosome function remains largely unknown. Here we show that a cluster of methylated nucleotides in domain IV of 25S rRNA is critical for integrity of the large ribosomal subunit. We identified the elusive cytosine-5 methyltransferase for C2278 in yeast as Rcm1 and found that a combined loss of cytosine-5 methylation at C2278 and ribose methylation at G2288 caused dramatic ribosome instability, resulting in loss of 60S ribosomal subunits. Structural and biochemical analyses revealed that this instability was caused by changes in the structure of 25S rRNA and a consequent loss of multiple ribosomal proteins from the large ribosomal subunit. Our data demonstrate that individual RNA modifications can strongly affect structure of large ribonucleoprotein complexes.

Keywords
RNA modification ribosomal RNA ribosome stability translation yeast
MeSH Terms
Methylation Methyltransferases/genetics,metabolism Protein Biosynthesis Protein Conformation RNA, Fungal/genetics RNA, Ribosomal/chemistry,genetics,metabolism RNA, Small Interfering/genetics Ribosomal Proteins/antagonists & inhibitors,genetics,metabolism Ribosomes/chemistry,genetics,metabolism Saccharomyces cerevisiae/genetics,growth & development,metabolism Saccharomyces cerevisiae Proteins/antagonists & inhibitors,chemistry,genetics,metabolism
Chemicals
RNA, Fungal RNA, Ribosomal RNA, Small Interfering RPL23A protein, S cerevisiae Ribosomal Proteins Saccharomyces cerevisiae Proteins RNA, ribosomal, 25S Methyltransferases
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Gigova Andriana
Biochemistry Center and Cluster of Excellence CellNetworks, University of Heidelberg, 69120 Heidelberg, Germany.
Duggimpudi Sujitha
Biochemistry Center and Cluster of Excellence CellNetworks, University of Heidelberg, 69120 Heidelberg, Germany.
Pollex Tim
Division of Epigenetics, DKFZ-ZMBH Alliance, German Cancer Research Center, 69120 Heidelberg, Germany.
Schaefer Matthias
Division of Epigenetics, DKFZ-ZMBH Alliance, German Cancer Research Center, 69120 Heidelberg, Germany.
Koš Martin
Biochemistry Center and Cluster of Excellence CellNetworks, University of Heidelberg, 69120 Heidelberg, Germany martin.kos@bzh.uni-heidelberg.de.
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Article Info
Journal
RNA (New York, N.Y.)
Abbr.
RNA
ISSN
1469-9001
DOI
10.1261/rna.043398.113
Email
martin.kos@bzh.uni-heidelberg.de
Published
2014-10-00
Epub
2014-00-14
Pages
1632-44
Language
English
Region
United States
NLM ID
9509184
PMCID
PMC4174444
Subset
IM
External Links
PubMed source DOI: 10.1261/rna.043398.113 PMC Full Text View journal details
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