Methylation at position 32 of tRNA catalyzed by TrmJ alters oxidative stress response in Pseudomonas aeruginosa
Name
Jaroensuk-2016-Methylation at position 32 of t.pdf
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4.63 MB
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Author(s)
Jaroensuk, Juthamas
Atichartpongkul, Sopapan
Wong, Yee Hwa
Liew, Chong Wai
McBee, Megan E.
Thongdee, Narumon
Prestwich, Erin G.
Mongkolsuk, Skorn
Lescar, Julien
Fuangthong, Mayuree
Date Issued
September 2016
Journal
Nucleic Acids Research
Publisher
Oxford University Press
Citation
Jaroensuk, Juthamas, et al. “ Methylation at Position 32 of tRNA Catalyzed by TrmJ Alters Oxidative Stress Response in Pseudomonas Aeruginosa.” Nucleic Acids Research 44, no. 22 (September 28, 2016): 10834–10848.
Version
Final published version
Abstract
Bacteria respond to environmental stresses using a variety of signaling and gene expression pathways, with translational mechanisms being the least well understood. Here, we identified a tRNA methyltransferase in Pseudomonas aeruginosa PA14, trmJ, which confers resistance to oxidative stress. Analysis of tRNA from a trmJ mutant revealed that TrmJ catalyzes formation of Cm, Um, and, unexpectedly, Am. Defined in vitro analyses revealed that tRNA[superscript Met(CAU)] and tRNA[superscript Trp(CCA)] are substrates for Cm formation, tRNA[superscript Gln(UUG)], tRNA[superscript Pro(UGG)], tRNA[superscript Pro(CGG)] and tRNA[superscript His(GUG)] for Um, and tRNA[superscript Pro(GGG)] for Am. tRNA[superscript Ser(UGA)], previously observed as a TrmJ substrate in Escherichia coli, was not modified by PA14 TrmJ. Position 32 was confirmed as the TrmJ target for Am in tRNA[superscriptPro(GGG)] and Um in tRNA[superscript Gln(UUG)] by mass spectrometric analysis. Crystal structures of the free catalytic N-terminal domain of TrmJ show a 2-fold symmetrical dimer with an active site located at the interface between the monomers and a flexible basic loop positioned to bind tRNA, with conformational changes upon binding of the SAM-analog sinefungin. The loss of TrmJ rendered PA14 sensitive to H2O2 exposure, with reduced expression of oxyR-recG, katB-ankB, and katE. These results reveal that TrmJ is a tRNA:Cm32/Um32/Am32 methyltransferase involved in translational fidelity and the oxidative stress response.
MIT Department
Massachusetts Institute of Technology. Department of Biological Engineering
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Creative Commons Attribution-NonCommercial 4.0 International
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DOI of Published Version
https://doi.org/10.1093/nar/gkw870
