Details
| Originalsprache | Englisch |
|---|---|
| Seiten (von - bis) | 1430-1437 |
| Seitenumfang | 8 |
| Fachzeitschrift | CHEMBIOCHEM |
| Jahrgang | 20 |
| Ausgabenummer | 11 |
| Frühes Online-Datum | 15 Jan. 2019 |
| Publikationsstatus | Veröffentlicht - 3 Juni 2019 |
Abstract
Transfer RNA (tRNA) modifications impact the structure and function of tRNAs, thus affecting the efficiency and fidelity of translation. In the opportunistic pathogen Pseudomonas aeruginosa translational regulation plays an important but less defined role in adaptation to changing environments. In this study, we have explored tRNA modifications in P. aeruginosa through LC-MS/MS approaches. Neutral loss scanning (NLS) demonstrated the potential to identify previously unknown modifications, whereas multiple reaction monitoring (MRM) was able to detect modifications with high specificity and sensitivity. In this study, the MRM-based external calibration method allowed for quantification of the four canonical and 32 modified ribonucleosides, out of which 21 tRNA modifications were quantified in the total tRNA pool of P. aeruginosa PA14. We also purified the single tRNA isoacceptors tRNA-ArgUCU, tRNA-LeuCAA, and tRNA-TrpCCA and determined their specific modification patterns, both qualitatively and quantitatively. Deeper insights into the nature and dynamics of tRNA modifications in P. aeruginosa should pave the way for further studies on post-transcriptional gene regulation as a relatively unexplored molecular mechanism of controlling bacterial pathogenicity and mode of growth.
ASJC Scopus Sachgebiete
- Biochemie, Genetik und Molekularbiologie (insg.)
- Biochemie
- Biochemie, Genetik und Molekularbiologie (insg.)
- Molekularmedizin
- Biochemie, Genetik und Molekularbiologie (insg.)
- Molekularbiologie
- Chemie (insg.)
- Organische Chemie
Zitieren
- Standard
- Harvard
- Apa
- Vancouver
- BibTex
- RIS
in: CHEMBIOCHEM, Jahrgang 20, Nr. 11, 03.06.2019, S. 1430-1437.
Publikation: Beitrag in Fachzeitschrift › Artikel › Forschung › Peer-Review
}
TY - JOUR
T1 - Identification and Quantification of (t)RNA Modifications in Pseudomonas aeruginosa by Liquid Chromatography–Tandem Mass Spectrometry
AU - Grobe, Svenja
AU - Doberenz, Sebastian
AU - Ferreira, Kevin
AU - Krueger, Jonas
AU - Brönstrup, Mark
AU - Kaever, Volkhard
AU - Häussler, Susanne
N1 - Funding Information: We gratefully thank Annette Garbe and Anna-Lena Hagemann for support with LC-MS/MS analysis, and Laura Santos for support with the resynthesis. V.K. and S.H. received funding from the German Research Foundation (DFG SPP 1879; speaker: Regine Hengge).
PY - 2019/6/3
Y1 - 2019/6/3
N2 - Transfer RNA (tRNA) modifications impact the structure and function of tRNAs, thus affecting the efficiency and fidelity of translation. In the opportunistic pathogen Pseudomonas aeruginosa translational regulation plays an important but less defined role in adaptation to changing environments. In this study, we have explored tRNA modifications in P. aeruginosa through LC-MS/MS approaches. Neutral loss scanning (NLS) demonstrated the potential to identify previously unknown modifications, whereas multiple reaction monitoring (MRM) was able to detect modifications with high specificity and sensitivity. In this study, the MRM-based external calibration method allowed for quantification of the four canonical and 32 modified ribonucleosides, out of which 21 tRNA modifications were quantified in the total tRNA pool of P. aeruginosa PA14. We also purified the single tRNA isoacceptors tRNA-ArgUCU, tRNA-LeuCAA, and tRNA-TrpCCA and determined their specific modification patterns, both qualitatively and quantitatively. Deeper insights into the nature and dynamics of tRNA modifications in P. aeruginosa should pave the way for further studies on post-transcriptional gene regulation as a relatively unexplored molecular mechanism of controlling bacterial pathogenicity and mode of growth.
AB - Transfer RNA (tRNA) modifications impact the structure and function of tRNAs, thus affecting the efficiency and fidelity of translation. In the opportunistic pathogen Pseudomonas aeruginosa translational regulation plays an important but less defined role in adaptation to changing environments. In this study, we have explored tRNA modifications in P. aeruginosa through LC-MS/MS approaches. Neutral loss scanning (NLS) demonstrated the potential to identify previously unknown modifications, whereas multiple reaction monitoring (MRM) was able to detect modifications with high specificity and sensitivity. In this study, the MRM-based external calibration method allowed for quantification of the four canonical and 32 modified ribonucleosides, out of which 21 tRNA modifications were quantified in the total tRNA pool of P. aeruginosa PA14. We also purified the single tRNA isoacceptors tRNA-ArgUCU, tRNA-LeuCAA, and tRNA-TrpCCA and determined their specific modification patterns, both qualitatively and quantitatively. Deeper insights into the nature and dynamics of tRNA modifications in P. aeruginosa should pave the way for further studies on post-transcriptional gene regulation as a relatively unexplored molecular mechanism of controlling bacterial pathogenicity and mode of growth.
KW - gene expression
KW - mass spectrometry
KW - Pseudomonas aeruginosa
KW - tRNA
KW - tRNA modifications
UR - http://www.scopus.com/inward/record.url?scp=85063905920&partnerID=8YFLogxK
U2 - 10.1002/cbic.201800741
DO - 10.1002/cbic.201800741
M3 - Article
C2 - 30644616
AN - SCOPUS:85063905920
VL - 20
SP - 1430
EP - 1437
JO - CHEMBIOCHEM
JF - CHEMBIOCHEM
SN - 1439-4227
IS - 11
ER -