The aminoglycoside resistance methyltransferase Sgm impedes RsmF methylation at an adjacent rRNA nucleotide in the ribosomal A site

Sonja Cubrilo, Fedora Babić, Stephen Douthwaite, Gordana Maravić Vlahovicek

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Resumé

Udgivelsesdato: Aug
OriginalsprogEngelsk
TidsskriftRNA: A publication of the RNA Society
Vol/bind15
Udgave nummer8
Sider (fra-til)1492-1497
Antal sider5
ISSN1355-8382
DOI
StatusUdgivet - 1. aug. 2009

Fingeraftryk

S-Adenosylmethionine
Microbial Drug Resistance
Protein Binding
Enzymes

Citer dette

@article{96e36ef0a2a311de9743000ea68e967b,
title = "The aminoglycoside resistance methyltransferase Sgm impedes RsmF methylation at an adjacent rRNA nucleotide in the ribosomal A site",
abstract = "Ribosome-targeting antibiotics block protein synthesis by binding at functionally important regions of the bacterial rRNA. Resistance is often conferred by addition of a methyl group at the antibiotic binding site within an rRNA region that is already highly modified with several nucleotide methylations. In bacterial rRNA, each methylation requires its own specific methyltransferase enzyme, and this raises the question as to how an extra methyltransferase conferring antibiotic resistance can be accommodated and how it can gain access to its nucleotide target within a short and functionally crowded stretch of the rRNA sequence. Here, we show that the Sgm methyltransferase confers resistance to 4,6-disubstituted deoxystreptamine aminoglycosides by introducing the 16S rRNA modification m(7)G1405 within the ribosomal A site. This region of Escherichia coli 16S rRNA already contains several methylated nucleotides including m(4)Cm1402 and m(5)C1407. Modification at m(5)C1407 by the methyltransferase RsmF is impeded as Sgm gains access to its adjacent G1405 target on the 30S ribosomal subunit. An Sgm mutant (G135A), which is impaired in S-adenosylmethionine binding and confers lower resistance, is less able to interfere with RsmF methylation on the 30S subunit. The two methylations at 16S rRNA nucleotide m(4)Cm1402 are unaffected by both the wild-type and the mutant versions of Sgm. The data indicate that interplay between resistance methyltransferases and the cell's own indigenous methyltransferases can play an important role in determining resistance levels.",
keywords = "Aminoglycosides, Bacterial Proteins, Base Sequence, Drug Resistance, Bacterial, Escherichia coli, Escherichia coli Proteins, Methylation, Methyltransferases, Mutagenesis, Site-Directed, Nucleic Acid Conformation, RNA, Bacterial, RNA, Ribosomal, RNA, Ribosomal, 16S, Recombinant Proteins, Ribosomes, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Substrate Specificity",
author = "Sonja Cubrilo and Fedora Babić and Stephen Douthwaite and Vlahovicek, {Gordana Maravić}",
year = "2009",
month = "8",
day = "1",
doi = "10.1261/rna.1618809",
language = "English",
volume = "15",
pages = "1492--1497",
journal = "R N A",
issn = "1355-8382",
publisher = "Cold Spring Harbor Laboratory Press",
number = "8",

}

The aminoglycoside resistance methyltransferase Sgm impedes RsmF methylation at an adjacent rRNA nucleotide in the ribosomal A site. / Cubrilo, Sonja; Babić, Fedora; Douthwaite, Stephen; Vlahovicek, Gordana Maravić.

I: RNA: A publication of the RNA Society, Bind 15, Nr. 8, 01.08.2009, s. 1492-1497.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - The aminoglycoside resistance methyltransferase Sgm impedes RsmF methylation at an adjacent rRNA nucleotide in the ribosomal A site

AU - Cubrilo, Sonja

AU - Babić, Fedora

AU - Douthwaite, Stephen

AU - Vlahovicek, Gordana Maravić

PY - 2009/8/1

Y1 - 2009/8/1

N2 - Ribosome-targeting antibiotics block protein synthesis by binding at functionally important regions of the bacterial rRNA. Resistance is often conferred by addition of a methyl group at the antibiotic binding site within an rRNA region that is already highly modified with several nucleotide methylations. In bacterial rRNA, each methylation requires its own specific methyltransferase enzyme, and this raises the question as to how an extra methyltransferase conferring antibiotic resistance can be accommodated and how it can gain access to its nucleotide target within a short and functionally crowded stretch of the rRNA sequence. Here, we show that the Sgm methyltransferase confers resistance to 4,6-disubstituted deoxystreptamine aminoglycosides by introducing the 16S rRNA modification m(7)G1405 within the ribosomal A site. This region of Escherichia coli 16S rRNA already contains several methylated nucleotides including m(4)Cm1402 and m(5)C1407. Modification at m(5)C1407 by the methyltransferase RsmF is impeded as Sgm gains access to its adjacent G1405 target on the 30S ribosomal subunit. An Sgm mutant (G135A), which is impaired in S-adenosylmethionine binding and confers lower resistance, is less able to interfere with RsmF methylation on the 30S subunit. The two methylations at 16S rRNA nucleotide m(4)Cm1402 are unaffected by both the wild-type and the mutant versions of Sgm. The data indicate that interplay between resistance methyltransferases and the cell's own indigenous methyltransferases can play an important role in determining resistance levels.

AB - Ribosome-targeting antibiotics block protein synthesis by binding at functionally important regions of the bacterial rRNA. Resistance is often conferred by addition of a methyl group at the antibiotic binding site within an rRNA region that is already highly modified with several nucleotide methylations. In bacterial rRNA, each methylation requires its own specific methyltransferase enzyme, and this raises the question as to how an extra methyltransferase conferring antibiotic resistance can be accommodated and how it can gain access to its nucleotide target within a short and functionally crowded stretch of the rRNA sequence. Here, we show that the Sgm methyltransferase confers resistance to 4,6-disubstituted deoxystreptamine aminoglycosides by introducing the 16S rRNA modification m(7)G1405 within the ribosomal A site. This region of Escherichia coli 16S rRNA already contains several methylated nucleotides including m(4)Cm1402 and m(5)C1407. Modification at m(5)C1407 by the methyltransferase RsmF is impeded as Sgm gains access to its adjacent G1405 target on the 30S ribosomal subunit. An Sgm mutant (G135A), which is impaired in S-adenosylmethionine binding and confers lower resistance, is less able to interfere with RsmF methylation on the 30S subunit. The two methylations at 16S rRNA nucleotide m(4)Cm1402 are unaffected by both the wild-type and the mutant versions of Sgm. The data indicate that interplay between resistance methyltransferases and the cell's own indigenous methyltransferases can play an important role in determining resistance levels.

KW - Aminoglycosides

KW - Bacterial Proteins

KW - Base Sequence

KW - Drug Resistance, Bacterial

KW - Escherichia coli

KW - Escherichia coli Proteins

KW - Methylation

KW - Methyltransferases

KW - Mutagenesis, Site-Directed

KW - Nucleic Acid Conformation

KW - RNA, Bacterial

KW - RNA, Ribosomal

KW - RNA, Ribosomal, 16S

KW - Recombinant Proteins

KW - Ribosomes

KW - Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

KW - Substrate Specificity

U2 - 10.1261/rna.1618809

DO - 10.1261/rna.1618809

M3 - Journal article

C2 - 19509304

VL - 15

SP - 1492

EP - 1497

JO - R N A

JF - R N A

SN - 1355-8382

IS - 8

ER -