The flavoprotein Mcap0476 (RlmFO) catalyzes m5U1939 modification in Mycoplasma capricolum 23S rRNA

Carole Lartigue, Anne Lebaudy, Alain Blanchard, Basma El Yacoubi, Simon Rose, Henri Grosjean, Stephen Roger Douthwaite

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Resumé

Efficient protein synthesis in all organisms requires the post-transcriptional methylation of specific ribosomal ribonucleic acid (rRNA) and transfer RNA (tRNA) nucleotides. The methylation reactions are almost invariably catalyzed by enzymes that use S-adenosylmethionine (AdoMet) as the methyl group donor. One noteworthy exception is seen in some bacteria, where the conserved tRNA methylation at m5U54 is added by the enzyme TrmFO using flavin adenine dinucleotide together with N5,N10-methylenetetrahydrofolate as the one-carbon donor. The minimalist bacterium Mycoplasma capricolum possesses two homologs of trmFO, but surprisingly lacks the m5U54 tRNA modification. We created single and dual deletions of the trmFO homologs using a novel synthetic biology approach. Subsequent analysis of the M. capricolum RNAs by mass spectrometry shows that the TrmFO homolog encoded by Mcap0476 specifically modifies m5U1939 in 23S rRNA, a conserved methylation catalyzed by AdoMet-dependent enzymes in all other characterized bacteria. The Mcap0476 methyltransferase (renamed RlmFO) represents the first folate-dependent flavoprotein seen to modify ribosomal RNA.

OriginalsprogEngelsk
TidsskriftNucleic Acids Research
Vol/bind42
Udgave nummer12
Sider (fra-til)8073-82
Antal sider10
ISSN0305-1048
DOI
StatusUdgivet - 2014

Fingeraftryk

Mycoplasma capricolum
Flavoproteins
Transfer RNA
RNA
Enzymes
S-Adenosylmethionine
Flavin-Adenine Dinucleotide
Ribosomal RNA
Proteins

Citer dette

Lartigue, Carole ; Lebaudy, Anne ; Blanchard, Alain ; El Yacoubi, Basma ; Rose, Simon ; Grosjean, Henri ; Douthwaite, Stephen Roger. / The flavoprotein Mcap0476 (RlmFO) catalyzes m5U1939 modification in Mycoplasma capricolum 23S rRNA. I: Nucleic Acids Research. 2014 ; Bind 42, Nr. 12. s. 8073-82.
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title = "The flavoprotein Mcap0476 (RlmFO) catalyzes m5U1939 modification in Mycoplasma capricolum 23S rRNA",
abstract = "Efficient protein synthesis in all organisms requires the post-transcriptional methylation of specific ribosomal ribonucleic acid (rRNA) and transfer RNA (tRNA) nucleotides. The methylation reactions are almost invariably catalyzed by enzymes that use S-adenosylmethionine (AdoMet) as the methyl group donor. One noteworthy exception is seen in some bacteria, where the conserved tRNA methylation at m5U54 is added by the enzyme TrmFO using flavin adenine dinucleotide together with N5,N10-methylenetetrahydrofolate as the one-carbon donor. The minimalist bacterium Mycoplasma capricolum possesses two homologs of trmFO, but surprisingly lacks the m5U54 tRNA modification. We created single and dual deletions of the trmFO homologs using a novel synthetic biology approach. Subsequent analysis of the M. capricolum RNAs by mass spectrometry shows that the TrmFO homolog encoded by Mcap0476 specifically modifies m5U1939 in 23S rRNA, a conserved methylation catalyzed by AdoMet-dependent enzymes in all other characterized bacteria. The Mcap0476 methyltransferase (renamed RlmFO) represents the first folate-dependent flavoprotein seen to modify ribosomal RNA.",
keywords = "Bacterial Proteins, Biocatalysis, Flavoproteins, Methylation, Methyltransferases, Mycoplasma capricolum, RNA, Ribosomal, 23S, RNA, Transfer, Uridine",
author = "Carole Lartigue and Anne Lebaudy and Alain Blanchard and {El Yacoubi}, Basma and Simon Rose and Henri Grosjean and Douthwaite, {Stephen Roger}",
note = "{\circledC} The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research.",
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The flavoprotein Mcap0476 (RlmFO) catalyzes m5U1939 modification in Mycoplasma capricolum 23S rRNA. / Lartigue, Carole; Lebaudy, Anne; Blanchard, Alain; El Yacoubi, Basma; Rose, Simon; Grosjean, Henri; Douthwaite, Stephen Roger.

I: Nucleic Acids Research, Bind 42, Nr. 12, 2014, s. 8073-82.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - The flavoprotein Mcap0476 (RlmFO) catalyzes m5U1939 modification in Mycoplasma capricolum 23S rRNA

AU - Lartigue, Carole

AU - Lebaudy, Anne

AU - Blanchard, Alain

AU - El Yacoubi, Basma

AU - Rose, Simon

AU - Grosjean, Henri

AU - Douthwaite, Stephen Roger

N1 - © The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research.

PY - 2014

Y1 - 2014

N2 - Efficient protein synthesis in all organisms requires the post-transcriptional methylation of specific ribosomal ribonucleic acid (rRNA) and transfer RNA (tRNA) nucleotides. The methylation reactions are almost invariably catalyzed by enzymes that use S-adenosylmethionine (AdoMet) as the methyl group donor. One noteworthy exception is seen in some bacteria, where the conserved tRNA methylation at m5U54 is added by the enzyme TrmFO using flavin adenine dinucleotide together with N5,N10-methylenetetrahydrofolate as the one-carbon donor. The minimalist bacterium Mycoplasma capricolum possesses two homologs of trmFO, but surprisingly lacks the m5U54 tRNA modification. We created single and dual deletions of the trmFO homologs using a novel synthetic biology approach. Subsequent analysis of the M. capricolum RNAs by mass spectrometry shows that the TrmFO homolog encoded by Mcap0476 specifically modifies m5U1939 in 23S rRNA, a conserved methylation catalyzed by AdoMet-dependent enzymes in all other characterized bacteria. The Mcap0476 methyltransferase (renamed RlmFO) represents the first folate-dependent flavoprotein seen to modify ribosomal RNA.

AB - Efficient protein synthesis in all organisms requires the post-transcriptional methylation of specific ribosomal ribonucleic acid (rRNA) and transfer RNA (tRNA) nucleotides. The methylation reactions are almost invariably catalyzed by enzymes that use S-adenosylmethionine (AdoMet) as the methyl group donor. One noteworthy exception is seen in some bacteria, where the conserved tRNA methylation at m5U54 is added by the enzyme TrmFO using flavin adenine dinucleotide together with N5,N10-methylenetetrahydrofolate as the one-carbon donor. The minimalist bacterium Mycoplasma capricolum possesses two homologs of trmFO, but surprisingly lacks the m5U54 tRNA modification. We created single and dual deletions of the trmFO homologs using a novel synthetic biology approach. Subsequent analysis of the M. capricolum RNAs by mass spectrometry shows that the TrmFO homolog encoded by Mcap0476 specifically modifies m5U1939 in 23S rRNA, a conserved methylation catalyzed by AdoMet-dependent enzymes in all other characterized bacteria. The Mcap0476 methyltransferase (renamed RlmFO) represents the first folate-dependent flavoprotein seen to modify ribosomal RNA.

KW - Bacterial Proteins

KW - Biocatalysis

KW - Flavoproteins

KW - Methylation

KW - Methyltransferases

KW - Mycoplasma capricolum

KW - RNA, Ribosomal, 23S

KW - RNA, Transfer

KW - Uridine

U2 - 10.1093/nar/gku518

DO - 10.1093/nar/gku518

M3 - Journal article

C2 - 24939895

VL - 42

SP - 8073

EP - 8082

JO - Nucleic Acids Research

JF - Nucleic Acids Research

SN - 0305-1048

IS - 12

ER -