Identification of modifications in microbial, native tRNA that suppress immunostimulatory activity

Publication type:
Journal article
Metadata:
Autoren
  • Stefanie Gehrig
  • Mariel-Esther Eberle
  • Flavia Botschen
  • Katharina Rimbach
  • Florian Eberle
  • Tatjana Eigenbrod
  • Steffen Kaiser
  • Walter M Holmes
  • Volker A Erdmann
  • Mathias Sprinzl
  • Guillaume Bec
  • Gerard Keith
  • Alexander H Dalpke
  • Mark Helm
Autoren-URL
https://www.webofscience.com/api/gateway?GWVersion=2&SrcApp=fis-test-1&SrcAuth=WosAPI&KeyUT=WOS:000301943200004&DestLinkType=FullRecord&DestApp=WOS_CPL
DOI
10.1084/jem.20111044
Externe Identifier
  • Clarivate Analytics Document Solution ID: 914IO
  • PubMed Identifier: 22312113
ISSN
0022-1007
Ausgabe der Veröffentlichung
2
Zeitschrift
JOURNAL OF EXPERIMENTAL MEDICINE
Paginierung
225 - 233
Datum der Veröffentlichung
2012
Status
Published
Titel
Identification of modifications in microbial, native tRNA that suppress immunostimulatory activity
Sub types
  • Article
Ausgabe der Zeitschrift
209

Data source: Web of Science (Lite)

Other metadata sources:
Abstract
<jats:p>Naturally occurring nucleotide modifications within RNA have been proposed to be structural determinants for innate immune recognition. We tested this hypothesis in the context of native nonself-RNAs. Isolated, fully modified native bacterial transfer RNAs (tRNAs) induced significant secretion of IFN-α from human peripheral blood mononuclear cells in a manner dependent on TLR7 and plasmacytoid dendritic cells. As a notable exception, tRNATyr from Escherichia coli was not immunostimulatory, as were all tested eukaryotic tRNAs. However, the unmodified, 5′-unphosphorylated in vitro transcript of tRNATyr induced IFN-α, thus revealing posttranscriptional modifications as a factor suppressing immunostimulation. Using a molecular surgery approach based on catalytic DNA, a panel of tRNATyr variants featuring differential modification patterns was examined. Out of seven modifications present in this tRNA, 2′-O-methylated Gm18 was identified as necessary and sufficient to suppress immunostimulation. Transplantation of this modification into the scaffold of yeast tRNAPhe also resulted in blocked immunostimulation. Moreover, an RNA preparation of an E. coli trmH mutant that lacks Gm18 2′-O-methyltransferase activity was significantly more stimulatory than the wild-type sample. The experiments identify the single methyl group on the 2′-oxygen of Gm18 as a natural modification in native tRNA that, beyond its primary structural role, has acquired a secondary function as an antagonist of TLR7.</jats:p>
Autoren
  • Stefanie Gehrig
  • Mariel-Esther Eberle
  • Flavia Botschen
  • Katharina Rimbach
  • Florian Eberle
  • Tatjana Eigenbrod
  • Steffen Kaiser
  • Walter M Holmes
  • Volker A Erdmann
  • Mathias Sprinzl
  • Guillaume Bec
  • Gérard Keith
  • Alexander H Dalpke
  • Mark Helm
DOI
10.1084/jem.20111044
eISSN
1540-9538
ISSN
0022-1007
Ausgabe der Veröffentlichung
2
Zeitschrift
Journal of Experimental Medicine
Sprache
en
Online publication date
2012
Paginierung
225 - 233
Datum der Veröffentlichung
2012
Status
Published
Herausgeber
Rockefeller University Press
Herausgeber URL
http://dx.doi.org/10.1084/jem.20111044
Datum der Datenerfassung
2023
Titel
Identification of modifications in microbial, native tRNA that suppress immunostimulatory activity
Ausgabe der Zeitschrift
209

Data source: Crossref

Abstract
Naturally occurring nucleotide modifications within RNA have been proposed to be structural determinants for innate immune recognition. We tested this hypothesis in the context of native nonself-RNAs. Isolated, fully modified native bacterial transfer RNAs (tRNAs) induced significant secretion of IFN-α from human peripheral blood mononuclear cells in a manner dependent on TLR7 and plasmacytoid dendritic cells. As a notable exception, tRNA(Tyr) from Escherichia coli was not immunostimulatory, as were all tested eukaryotic tRNAs. However, the unmodified, 5'-unphosphorylated in vitro transcript of tRNA(Tyr) induced IFN-α, thus revealing posttranscriptional modifications as a factor suppressing immunostimulation. Using a molecular surgery approach based on catalytic DNA, a panel of tRNA(Tyr) variants featuring differential modification patterns was examined. Out of seven modifications present in this tRNA, 2'-O-methylated G(m)18 was identified as necessary and sufficient to suppress immunostimulation. Transplantation of this modification into the scaffold of yeast tRNA(Phe) also resulted in blocked immunostimulation. Moreover, an RNA preparation of an E. coli trmH mutant that lacks G(m)18 2'-O-methyltransferase activity was significantly more stimulatory than the wild-type sample. The experiments identify the single methyl group on the 2'-oxygen of G(m)18 as a natural modification in native tRNA that, beyond its primary structural role, has acquired a secondary function as an antagonist of TLR7.
Addresses
  • Department of Chemistry, Institute of Pharmacy and Molecular Biotechnology, Heidelberg University, 69117 Heidelberg, Germany.
Autoren
  • Stefanie Gehrig
  • Mariel-Esther Eberle
  • Flavia Botschen
  • Katharina Rimbach
  • Florian Eberle
  • Tatjana Eigenbrod
  • Steffen Kaiser
  • Walter M Holmes
  • Volker A Erdmann
  • Mathias Sprinzl
  • Guillaume Bec
  • Gérard Keith
  • Alexander H Dalpke
  • Mark Helm
DOI
10.1084/jem.20111044
eISSN
1540-9538
Externe Identifier
  • PubMed Identifier: 22312113
  • PubMed Central ID: PMC3280868
Open access
true
ISSN
0022-1007
Ausgabe der Veröffentlichung
2
Zeitschrift
The Journal of experimental medicine
Schlüsselwörter
  • Dendritic Cells
  • Leukocytes, Mononuclear
  • Humans
  • Escherichia coli
  • tRNA Methyltransferases
  • RNA, Transfer, Amino Acyl
  • Interferon-alpha
  • DNA Primers
  • Enzyme-Linked Immunosorbent Assay
  • Immunization
  • Electrophoresis, Polyacrylamide Gel
  • RNA Processing, Post-Transcriptional
  • Phosphorylation
  • Toll-Like Receptor 7
  • Immunity, Innate
Sprache
eng
Medium
Print-Electronic
Online publication date
2012
Open access status
Open Access
Paginierung
225 - 233
Datum der Veröffentlichung
2012
Status
Published
Publisher licence
CC BY-NC-SA
Datum der Datenerfassung
2012
Titel
Identification of modifications in microbial, native tRNA that suppress immunostimulatory activity.
Sub types
  • Research Support, Non-U.S. Gov't
  • research-article
  • Journal Article
Ausgabe der Zeitschrift
209

Files

https://rupress.org/jem/article-pdf/209/2/225/539887/jem_20111044.pdf https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22312113/pdf/?tool=EBI https://europepmc.org/articles/PMC3280868?pdf=render

Data source: Europe PubMed Central

Abstract
Naturally occurring nucleotide modifications within RNA have been proposed to be structural determinants for innate immune recognition. We tested this hypothesis in the context of native nonself-RNAs. Isolated, fully modified native bacterial transfer RNAs (tRNAs) induced significant secretion of IFN-α from human peripheral blood mononuclear cells in a manner dependent on TLR7 and plasmacytoid dendritic cells. As a notable exception, tRNA(Tyr) from Escherichia coli was not immunostimulatory, as were all tested eukaryotic tRNAs. However, the unmodified, 5'-unphosphorylated in vitro transcript of tRNA(Tyr) induced IFN-α, thus revealing posttranscriptional modifications as a factor suppressing immunostimulation. Using a molecular surgery approach based on catalytic DNA, a panel of tRNA(Tyr) variants featuring differential modification patterns was examined. Out of seven modifications present in this tRNA, 2'-O-methylated G(m)18 was identified as necessary and sufficient to suppress immunostimulation. Transplantation of this modification into the scaffold of yeast tRNA(Phe) also resulted in blocked immunostimulation. Moreover, an RNA preparation of an E. coli trmH mutant that lacks G(m)18 2'-O-methyltransferase activity was significantly more stimulatory than the wild-type sample. The experiments identify the single methyl group on the 2'-oxygen of G(m)18 as a natural modification in native tRNA that, beyond its primary structural role, has acquired a secondary function as an antagonist of TLR7.
Autoren
  • Stefanie Gehrig
  • Mariel-Esther Eberle
  • Flavia Botschen
  • Katharina Rimbach
  • Florian Eberle
  • Tatjana Eigenbrod
  • Steffen Kaiser
  • Walter M Holmes
  • Volker A Erdmann
  • Mathias Sprinzl
  • Guillaume Bec
  • Gérard Keith
  • Alexander H Dalpke
  • Mark Helm
Autoren-URL
https://www.ncbi.nlm.nih.gov/pubmed/22312113
DOI
10.1084/jem.20111044
eISSN
1540-9538
Externe Identifier
  • PubMed Central ID: PMC3280868
Ausgabe der Veröffentlichung
2
Zeitschrift
J Exp Med
Schlüsselwörter
  • DNA Primers
  • Dendritic Cells
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme-Linked Immunosorbent Assay
  • Escherichia coli
  • Humans
  • Immunity, Innate
  • Immunization
  • Interferon-alpha
  • Leukocytes, Mononuclear
  • Phosphorylation
  • RNA Processing, Post-Transcriptional
  • RNA, Transfer, Amino Acyl
  • Toll-Like Receptor 7
  • tRNA Methyltransferases
Sprache
eng
Country
United States
Paginierung
225 - 233
PII
jem.20111044
Datum der Veröffentlichung
2012
Status
Published
Datum, an dem der Datensatz öffentlich gemacht wurde
2012
Titel
Identification of modifications in microbial, native tRNA that suppress immunostimulatory activity.
Sub types
  • Journal Article
  • Research Support, Non-U.S. Gov't
Ausgabe der Zeitschrift
209

Data source: PubMed

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