Loading…
Identifying the methyltransferases for m super(5)U747 and m super(5)U1939 in 23S rRNA using MALDI mass spectrometry
There are three sites of m super(5)U modification in Escherichia coli stable RNAs: one at the invariant tRNA position U54 and two in 23S rRNA at the phylogenetically conserved positions U747 and U1939. Each of these sites is modified by its own methyltransferase, and the tRNA methyltransferase, TrmA...
Saved in:
Published in: | Nucleic acids research 2003-01, Vol.31 (16), p.4738-4746 |
---|---|
Main Authors: | , , , |
Format: | Article |
Language: | English |
Online Access: | Get full text |
Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
cited_by | |
---|---|
cites | |
container_end_page | 4746 |
container_issue | 16 |
container_start_page | 4738 |
container_title | Nucleic acids research |
container_volume | 31 |
creator | Madsen, C T Mengel-Joergensen, J Kirpekar, F Douthwaite, S |
description | There are three sites of m super(5)U modification in Escherichia coli stable RNAs: one at the invariant tRNA position U54 and two in 23S rRNA at the phylogenetically conserved positions U747 and U1939. Each of these sites is modified by its own methyltransferase, and the tRNA methyltransferase, TrmA, is well-characterised. Two open reading frames, YbjF and YgcA, are approximately 30% identical to TrmA, and here we determine the functions of these candidate methyltransferases using MALDI mass spectrometry. A purified recombinant version of YgcA retains its activity and specificity, and methylates U1939 in an RNA transcript in vitro. We were unable to generate a recombinant version of YbjF that retained in vitro activity, so the function of this enzyme was defined in vivo by engineering a ybjF knockout strain. Comparison of the methylation patterns in 23S rRNAs from YbjF super(+) and YbjF strains showed that the latter differed only in the lack of the m super(5)U747 modification. With this report, the functions of all the E.coli m super(5)U RNA methyltransferases are identified, and a more appropriate designation for YbjF would be RumB (RNA uridine methyltransferases B), in line with the recent nomenclature change for YgcA (now RumA). |
format | article |
fullrecord | <record><control><sourceid>proquest</sourceid><recordid>TN_cdi_proquest_miscellaneous_18804364</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>18804364</sourcerecordid><originalsourceid>FETCH-proquest_miscellaneous_188043643</originalsourceid><addsrcrecordid>eNqNjLsKwjAUQDMoWB__cCfRQUhMattRfKCgDj7mEvTWVtq05qZD_14FB0enA4fDaTGPS-5PBFdhh3WJHpwLJXzlMdre0LgsaTJzB5ciFOjSJndWG0rQakKCpLRQANUV2pE_vgQqAG1uv0pEMoLMwFSewB4Pc6jp89vPd8stFJoIqMKrs-X7bps-ayc6Jxx82WPD9eq82EwqWz5rJBcXGV0xz7XBsqZYhCFXcqbk3-ELf3pLXQ</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>18804364</pqid></control><display><type>article</type><title>Identifying the methyltransferases for m super(5)U747 and m super(5)U1939 in 23S rRNA using MALDI mass spectrometry</title><source>Oxford Journals website</source><source>PubMed Central</source><creator>Madsen, C T ; Mengel-Joergensen, J ; Kirpekar, F ; Douthwaite, S</creator><creatorcontrib>Madsen, C T ; Mengel-Joergensen, J ; Kirpekar, F ; Douthwaite, S</creatorcontrib><description>There are three sites of m super(5)U modification in Escherichia coli stable RNAs: one at the invariant tRNA position U54 and two in 23S rRNA at the phylogenetically conserved positions U747 and U1939. Each of these sites is modified by its own methyltransferase, and the tRNA methyltransferase, TrmA, is well-characterised. Two open reading frames, YbjF and YgcA, are approximately 30% identical to TrmA, and here we determine the functions of these candidate methyltransferases using MALDI mass spectrometry. A purified recombinant version of YgcA retains its activity and specificity, and methylates U1939 in an RNA transcript in vitro. We were unable to generate a recombinant version of YbjF that retained in vitro activity, so the function of this enzyme was defined in vivo by engineering a ybjF knockout strain. Comparison of the methylation patterns in 23S rRNAs from YbjF super(+) and YbjF strains showed that the latter differed only in the lack of the m super(5)U747 modification. With this report, the functions of all the E.coli m super(5)U RNA methyltransferases are identified, and a more appropriate designation for YbjF would be RumB (RNA uridine methyltransferases B), in line with the recent nomenclature change for YgcA (now RumA).</description><identifier>ISSN: 0305-1048</identifier><language>eng</language><ispartof>Nucleic acids research, 2003-01, Vol.31 (16), p.4738-4746</ispartof><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784</link.rule.ids></links><search><creatorcontrib>Madsen, C T</creatorcontrib><creatorcontrib>Mengel-Joergensen, J</creatorcontrib><creatorcontrib>Kirpekar, F</creatorcontrib><creatorcontrib>Douthwaite, S</creatorcontrib><title>Identifying the methyltransferases for m super(5)U747 and m super(5)U1939 in 23S rRNA using MALDI mass spectrometry</title><title>Nucleic acids research</title><description>There are three sites of m super(5)U modification in Escherichia coli stable RNAs: one at the invariant tRNA position U54 and two in 23S rRNA at the phylogenetically conserved positions U747 and U1939. Each of these sites is modified by its own methyltransferase, and the tRNA methyltransferase, TrmA, is well-characterised. Two open reading frames, YbjF and YgcA, are approximately 30% identical to TrmA, and here we determine the functions of these candidate methyltransferases using MALDI mass spectrometry. A purified recombinant version of YgcA retains its activity and specificity, and methylates U1939 in an RNA transcript in vitro. We were unable to generate a recombinant version of YbjF that retained in vitro activity, so the function of this enzyme was defined in vivo by engineering a ybjF knockout strain. Comparison of the methylation patterns in 23S rRNAs from YbjF super(+) and YbjF strains showed that the latter differed only in the lack of the m super(5)U747 modification. With this report, the functions of all the E.coli m super(5)U RNA methyltransferases are identified, and a more appropriate designation for YbjF would be RumB (RNA uridine methyltransferases B), in line with the recent nomenclature change for YgcA (now RumA).</description><issn>0305-1048</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2003</creationdate><recordtype>article</recordtype><recordid>eNqNjLsKwjAUQDMoWB__cCfRQUhMattRfKCgDj7mEvTWVtq05qZD_14FB0enA4fDaTGPS-5PBFdhh3WJHpwLJXzlMdre0LgsaTJzB5ciFOjSJndWG0rQakKCpLRQANUV2pE_vgQqAG1uv0pEMoLMwFSewB4Pc6jp89vPd8stFJoIqMKrs-X7bps-ayc6Jxx82WPD9eq82EwqWz5rJBcXGV0xz7XBsqZYhCFXcqbk3-ELf3pLXQ</recordid><startdate>20030101</startdate><enddate>20030101</enddate><creator>Madsen, C T</creator><creator>Mengel-Joergensen, J</creator><creator>Kirpekar, F</creator><creator>Douthwaite, S</creator><scope>7QL</scope><scope>7TM</scope><scope>C1K</scope></search><sort><creationdate>20030101</creationdate><title>Identifying the methyltransferases for m super(5)U747 and m super(5)U1939 in 23S rRNA using MALDI mass spectrometry</title><author>Madsen, C T ; Mengel-Joergensen, J ; Kirpekar, F ; Douthwaite, S</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-proquest_miscellaneous_188043643</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2003</creationdate><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Madsen, C T</creatorcontrib><creatorcontrib>Mengel-Joergensen, J</creatorcontrib><creatorcontrib>Kirpekar, F</creatorcontrib><creatorcontrib>Douthwaite, S</creatorcontrib><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Nucleic Acids Abstracts</collection><collection>Environmental Sciences and Pollution Management</collection><jtitle>Nucleic acids research</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Madsen, C T</au><au>Mengel-Joergensen, J</au><au>Kirpekar, F</au><au>Douthwaite, S</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Identifying the methyltransferases for m super(5)U747 and m super(5)U1939 in 23S rRNA using MALDI mass spectrometry</atitle><jtitle>Nucleic acids research</jtitle><date>2003-01-01</date><risdate>2003</risdate><volume>31</volume><issue>16</issue><spage>4738</spage><epage>4746</epage><pages>4738-4746</pages><issn>0305-1048</issn><abstract>There are three sites of m super(5)U modification in Escherichia coli stable RNAs: one at the invariant tRNA position U54 and two in 23S rRNA at the phylogenetically conserved positions U747 and U1939. Each of these sites is modified by its own methyltransferase, and the tRNA methyltransferase, TrmA, is well-characterised. Two open reading frames, YbjF and YgcA, are approximately 30% identical to TrmA, and here we determine the functions of these candidate methyltransferases using MALDI mass spectrometry. A purified recombinant version of YgcA retains its activity and specificity, and methylates U1939 in an RNA transcript in vitro. We were unable to generate a recombinant version of YbjF that retained in vitro activity, so the function of this enzyme was defined in vivo by engineering a ybjF knockout strain. Comparison of the methylation patterns in 23S rRNAs from YbjF super(+) and YbjF strains showed that the latter differed only in the lack of the m super(5)U747 modification. With this report, the functions of all the E.coli m super(5)U RNA methyltransferases are identified, and a more appropriate designation for YbjF would be RumB (RNA uridine methyltransferases B), in line with the recent nomenclature change for YgcA (now RumA).</abstract></addata></record> |
fulltext | fulltext |
identifier | ISSN: 0305-1048 |
ispartof | Nucleic acids research, 2003-01, Vol.31 (16), p.4738-4746 |
issn | 0305-1048 |
language | eng |
recordid | cdi_proquest_miscellaneous_18804364 |
source | Oxford Journals website; PubMed Central |
title | Identifying the methyltransferases for m super(5)U747 and m super(5)U1939 in 23S rRNA using MALDI mass spectrometry |
url | http://sfxeu10.hosted.exlibrisgroup.com/loughborough?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-24T17%3A48%3A40IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Identifying%20the%20methyltransferases%20for%20m%20super(5)U747%20and%20m%20super(5)U1939%20in%2023S%20rRNA%20using%20MALDI%20mass%20spectrometry&rft.jtitle=Nucleic%20acids%20research&rft.au=Madsen,%20C%20T&rft.date=2003-01-01&rft.volume=31&rft.issue=16&rft.spage=4738&rft.epage=4746&rft.pages=4738-4746&rft.issn=0305-1048&rft_id=info:doi/&rft_dat=%3Cproquest%3E18804364%3C/proquest%3E%3Cgrp_id%3Ecdi_FETCH-proquest_miscellaneous_188043643%3C/grp_id%3E%3Coa%3E%3C/oa%3E%3Curl%3E%3C/url%3E&rft_id=info:oai/&rft_pqid=18804364&rft_id=info:pmid/&rfr_iscdi=true |