Partial Methylation at Am100 in 18S rRNA of Baker's Yeast Reveals Ribosome Heterogeneity on the Level of Eukaryotic rRNA Modification

Autoren

Markus Buchhaupt
Sunny Sharma
Stefanie Kellner
Stefanie Oswald
Melanie Paetzold
Christian Peifer
Peter Watzinger
Jens Schrader
Mark Helm
Karl-Dieter Entian

DOI

10.1371/journal.pone.0089640

Editoren

Thomas Preiss

eISSN

1932-6203

Ausgabe der Veröffentlichung

2

Zeitschrift

PLoS ONE

Sprache

en

Online publication date

2014

Paginierung

e89640 - e89640

Status

Published online

Herausgeber

Public Library of Science (PLoS)

Herausgeber URL

http://dx.doi.org/10.1371/journal.pone.0089640

Datum der Datenerfassung

2022

Titel

Partial Methylation at Am100 in 18S rRNA of Baker's Yeast Reveals Ribosome Heterogeneity on the Level of Eukaryotic rRNA Modification

Ausgabe der Zeitschrift

9

Datenquelle: Crossref

Abstract

Ribosome heterogeneity is of increasing biological significance and several examples have been described for multicellular and single cells organisms. In here we show for the first time a variation in ribose methylation within the 18S rRNA of Saccharomyces cerevisiae. Using RNA-cleaving DNAzymes, we could specifically demonstrate that a significant amount of S. cerevisiae ribosomes are not methylated at 2'-O-ribose of A100 residue in the 18S rRNA. Furthermore, using LC-UV-MS/MS of a respective 18S rRNA fragment, we could not only corroborate the partial methylation at A100, but could also quantify the methylated versus non-methylated A100 residue. Here, we exhibit that only 68% of A100 in the 18S rRNA of S.cerevisiae are methylated at 2'-O ribose sugar. Polysomes also contain a similar heterogeneity for methylated Am100, which shows that 40S ribosome subunits with and without Am100 participate in translation. Introduction of a multicopy plasmid containing the corresponding methylation guide snoRNA gene SNR51 led to an increased A100 methylation, suggesting the cellular snR51 level to limit the extent of this modification. Partial rRNA modification demonstrates a new level of ribosome heterogeneity in eukaryotic cells that might have substantial impact on regulation and fine-tuning of the translation process.

Addresses

DECHEMA Research Institute, Biochemical Engineering, Frankfurt am Main, Germany.

Autoren

Markus Buchhaupt
Sunny Sharma
Stefanie Kellner
Stefanie Oswald
Melanie Paetzold
Christian Peifer
Peter Watzinger
Jens Schrader
Mark Helm
Karl-Dieter Entian

DOI

10.1371/journal.pone.0089640

eISSN

1932-6203

Externe Identifier

PubMed Identifier: 24586927
PubMed Central ID: PMC3938493

Open access

true

ISSN

1932-6203

Ausgabe der Veröffentlichung

2

Zeitschrift

PloS one

Schlüsselwörter

Ribosomes
Saccharomyces cerevisiae
RNA, Ribosomal
Methylation

Sprache

eng

Medium

Electronic-eCollection

Online publication date

2014

Open access status

Open Access

Paginierung

e89640

Datum der Veröffentlichung

2014

Status

Published

Publisher licence

CC BY

Datum der Datenerfassung

2014

Titel

Partial methylation at Am100 in 18S rRNA of baker's yeast reveals ribosome heterogeneity on the level of eukaryotic rRNA modification.

Sub types

research-article
Journal Article

Ausgabe der Zeitschrift

9

Files

https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0089640&type=printable https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24586927/pdf/?tool=EBI https://europepmc.org/articles/PMC3938493?pdf=render

Datenquelle: Europe PubMed Central

Abstract

Ribosome heterogeneity is of increasing biological significance and several examples have been described for multicellular and single cells organisms. In here we show for the first time a variation in ribose methylation within the 18S rRNA of Saccharomyces cerevisiae. Using RNA-cleaving DNAzymes, we could specifically demonstrate that a significant amount of S. cerevisiae ribosomes are not methylated at 2'-O-ribose of A100 residue in the 18S rRNA. Furthermore, using LC-UV-MS/MS of a respective 18S rRNA fragment, we could not only corroborate the partial methylation at A100, but could also quantify the methylated versus non-methylated A100 residue. Here, we exhibit that only 68% of A100 in the 18S rRNA of S.cerevisiae are methylated at 2'-O ribose sugar. Polysomes also contain a similar heterogeneity for methylated Am100, which shows that 40S ribosome subunits with and without Am100 participate in translation. Introduction of a multicopy plasmid containing the corresponding methylation guide snoRNA gene SNR51 led to an increased A100 methylation, suggesting the cellular snR51 level to limit the extent of this modification. Partial rRNA modification demonstrates a new level of ribosome heterogeneity in eukaryotic cells that might have substantial impact on regulation and fine-tuning of the translation process.

Date of acceptance

2014

Autoren

Markus Buchhaupt
Sunny Sharma
Stefanie Kellner
Stefanie Oswald
Melanie Paetzold
Christian Peifer
Peter Watzinger
Jens Schrader
Mark Helm
Karl-Dieter Entian

Autoren-URL

https://www.ncbi.nlm.nih.gov/pubmed/24586927

DOI

10.1371/journal.pone.0089640

eISSN

1932-6203

Externe Identifier

PubMed Central ID: PMC3938493

Ausgabe der Veröffentlichung

2

Zeitschrift

PLoS One

Schlüsselwörter

Methylation
RNA, Ribosomal
Ribosomes
Saccharomyces cerevisiae

Sprache

eng

Country

United States

Paginierung

e89640

PII

PONE-D-13-01744

Datum der Veröffentlichung

2014

Status

Published online

Datum, an dem der Datensatz öffentlich gemacht wurde

2014

Titel

Partial methylation at Am100 in 18S rRNA of baker's yeast reveals ribosome heterogeneity on the level of eukaryotic rRNA modification.

Sub types

Journal Article

Ausgabe der Zeitschrift

9

Datenquelle: PubMed

Partial Methylation at Am100 in 18S rRNA of Baker's Yeast Reveals Ribosome Heterogeneity on the Level of Eukaryotic rRNA Modification

Files

Werkzeuge