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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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. |
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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.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0089640</identifier><identifier>PMID: 24586927</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Acids ; Adenosine ; Baking yeast ; Biochemistry ; Biology ; Cancer ; DNA methylation ; E coli ; Escherichia coli ; Genetic engineering ; Heterogeneity ; Methylation ; Monosaccharides ; Mutation ; Pharmacy ; Physiology ; Polyribosomes ; Proteins ; Ribonucleic acid ; Ribose ; Ribosomes ; Ribosomes - metabolism ; RNA ; RNA modification ; RNA, Ribosomal - genetics ; rRNA 18S ; Saccharomyces cerevisiae ; Saccharomyces cerevisiae - genetics ; snoRNA ; Sugar ; Translation ; Yeast</subject><ispartof>PloS one, 2014-02, Vol.9 (2), p.e89640-e89640</ispartof><rights>COPYRIGHT 2014 Public Library of Science</rights><rights>2014 Buchhaupt et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2014 Buchhaupt et al 2014 Buchhaupt et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c758t-4c4b5a4388832d1e30f36adb6f33942912f9401da7461949478f4ee27787946b3</citedby><cites>FETCH-LOGICAL-c758t-4c4b5a4388832d1e30f36adb6f33942912f9401da7461949478f4ee27787946b3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3938493/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3938493/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,864,885,2102,2928,23866,27924,27925,53791,53793,79600,79601</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/24586927$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Preiss, Thomas</contributor><creatorcontrib>Buchhaupt, Markus</creatorcontrib><creatorcontrib>Sharma, Sunny</creatorcontrib><creatorcontrib>Kellner, Stefanie</creatorcontrib><creatorcontrib>Oswald, Stefanie</creatorcontrib><creatorcontrib>Paetzold, Melanie</creatorcontrib><creatorcontrib>Peifer, Christian</creatorcontrib><creatorcontrib>Watzinger, Peter</creatorcontrib><creatorcontrib>Schrader, Jens</creatorcontrib><creatorcontrib>Helm, Mark</creatorcontrib><creatorcontrib>Entian, Karl-Dieter</creatorcontrib><title>Partial methylation at Am100 in 18S rRNA of baker's yeast reveals ribosome heterogeneity on the level of eukaryotic rRNA modification</title><title>PloS one</title><addtitle>PLoS One</addtitle><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.</description><subject>Acids</subject><subject>Adenosine</subject><subject>Baking yeast</subject><subject>Biochemistry</subject><subject>Biology</subject><subject>Cancer</subject><subject>DNA methylation</subject><subject>E coli</subject><subject>Escherichia coli</subject><subject>Genetic engineering</subject><subject>Heterogeneity</subject><subject>Methylation</subject><subject>Monosaccharides</subject><subject>Mutation</subject><subject>Pharmacy</subject><subject>Physiology</subject><subject>Polyribosomes</subject><subject>Proteins</subject><subject>Ribonucleic acid</subject><subject>Ribose</subject><subject>Ribosomes</subject><subject>Ribosomes - metabolism</subject><subject>RNA</subject><subject>RNA modification</subject><subject>RNA, Ribosomal - genetics</subject><subject>rRNA 18S</subject><subject>Saccharomyces cerevisiae</subject><subject>Saccharomyces cerevisiae - 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Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>PloS one</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Buchhaupt, Markus</au><au>Sharma, Sunny</au><au>Kellner, Stefanie</au><au>Oswald, Stefanie</au><au>Paetzold, Melanie</au><au>Peifer, Christian</au><au>Watzinger, Peter</au><au>Schrader, Jens</au><au>Helm, Mark</au><au>Entian, Karl-Dieter</au><au>Preiss, Thomas</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Partial methylation at Am100 in 18S rRNA of baker's yeast reveals ribosome heterogeneity on the level of eukaryotic rRNA modification</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2014-02-28</date><risdate>2014</risdate><volume>9</volume><issue>2</issue><spage>e89640</spage><epage>e89640</epage><pages>e89640-e89640</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><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.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>24586927</pmid><doi>10.1371/journal.pone.0089640</doi><tpages>e89640</tpages><oa>free_for_read</oa></addata></record> |
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subjects | Acids Adenosine Baking yeast Biochemistry Biology Cancer DNA methylation E coli Escherichia coli Genetic engineering Heterogeneity Methylation Monosaccharides Mutation Pharmacy Physiology Polyribosomes Proteins Ribonucleic acid Ribose Ribosomes Ribosomes - metabolism RNA RNA modification RNA, Ribosomal - genetics rRNA 18S Saccharomyces cerevisiae Saccharomyces cerevisiae - genetics snoRNA Sugar Translation Yeast |
title | Partial methylation at Am100 in 18S rRNA of baker's yeast reveals ribosome heterogeneity on the level of eukaryotic rRNA modification |
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