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

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...

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Autores principales: Markus Buchhaupt, Sunny Sharma, Stefanie Kellner, Stefanie Oswald, Melanie Paetzold, Christian Peifer, Peter Watzinger, Jens Schrader, Mark Helm, Karl-Dieter Entian
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spelling oai:doaj.org-article:176343c75ac94c268a08a627ead16d2c2021-11-18T08:30:31ZPartial methylation at Am100 in 18S rRNA of baker's yeast reveals ribosome heterogeneity on the level of eukaryotic rRNA modification.1932-620310.1371/journal.pone.0089640https://doaj.org/article/176343c75ac94c268a08a627ead16d2c2014-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24586927/?tool=EBIhttps://doaj.org/toc/1932-6203Ribosome 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.Markus BuchhauptSunny SharmaStefanie KellnerStefanie OswaldMelanie PaetzoldChristian PeiferPeter WatzingerJens SchraderMark HelmKarl-Dieter EntianPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 9, Iss 2, p e89640 (2014)
institution DOAJ
collection DOAJ
language EN
topic Medicine
R
Science
Q
spellingShingle Medicine
R
Science
Q
Markus Buchhaupt
Sunny Sharma
Stefanie Kellner
Stefanie Oswald
Melanie Paetzold
Christian Peifer
Peter Watzinger
Jens Schrader
Mark Helm
Karl-Dieter Entian
Partial methylation at Am100 in 18S rRNA of baker's yeast reveals ribosome heterogeneity on the level of eukaryotic rRNA modification.
description 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.
format article
author Markus Buchhaupt
Sunny Sharma
Stefanie Kellner
Stefanie Oswald
Melanie Paetzold
Christian Peifer
Peter Watzinger
Jens Schrader
Mark Helm
Karl-Dieter Entian
author_facet Markus Buchhaupt
Sunny Sharma
Stefanie Kellner
Stefanie Oswald
Melanie Paetzold
Christian Peifer
Peter Watzinger
Jens Schrader
Mark Helm
Karl-Dieter Entian
author_sort Markus Buchhaupt
title Partial methylation at Am100 in 18S rRNA of baker's yeast reveals ribosome heterogeneity on the level of eukaryotic rRNA modification.
title_short Partial methylation at Am100 in 18S rRNA of baker's yeast reveals ribosome heterogeneity on the level of eukaryotic rRNA modification.
title_full Partial methylation at Am100 in 18S rRNA of baker's yeast reveals ribosome heterogeneity on the level of eukaryotic rRNA modification.
title_fullStr Partial methylation at Am100 in 18S rRNA of baker's yeast reveals ribosome heterogeneity on the level of eukaryotic rRNA modification.
title_full_unstemmed Partial methylation at Am100 in 18S rRNA of baker's yeast reveals ribosome heterogeneity on the level of eukaryotic rRNA modification.
title_sort partial methylation at am100 in 18s rrna of baker's yeast reveals ribosome heterogeneity on the level of eukaryotic rrna modification.
publisher Public Library of Science (PLoS)
publishDate 2014
url https://doaj.org/article/176343c75ac94c268a08a627ead16d2c
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