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Repisalud author Prezioso, Samantha M author Duong, Duc M author Kuiper, Emily G author Deng, Qiudong author Alberti, Sebastian author Conn, Graeme L author Goldberg, Joanna B 2024-09-10T13:09:47Z 2024-09-10T13:09:47Z 2019-03-05 Prezioso SM, Duong DM, Kuiper EG, Deng Q, Alberti S, Conn GL, et al. Trimethylation of Elongation Factor-Tu by the Dual Thermoregulated Methyltransferase EftM Does Not Impact Its Canonical Function in Translation. Sci Rep. 2019 Mar 05;9:3553. 10.1038/s41598-019-39331-x 2045-2322 Scientific Reports http://hdl.handle.net/20.500.13003/17595 30837495 L626687782 2-s2.0-85062586175 https://hdl.handle.net/20.500.12105/22776 460381600126 The Pseudomonas aeruginosa methyltransferase EftM trimethylates elongation factor-Tu (EF-Tu) on lysine 5 to form a post-translational modification important for initial bacterial adherence to host epithelial cells. EftM methyltransferase activity is directly temperature regulated. The protein stability of EftM is tuned with a melting temperature (T-m) around 37 degrees C such that the enzyme is stable and active at 25 degrees C, but is completely inactivated by protein unfolding at higher temperatures. This leads to higher observable levels of EF-Tu trimethylation at the lower temperature. Here we report an additional layer of thermoregulation resulting in lower eftM mRNA transcript level at 37 degrees C compared to 25 degrees C and show that this regulation occurs at the level of transcription initiation. To begin to define the impact of this system on P. aeruginosa physiology, we demonstrate that EF-Tu is the only observable substrate for EftM. Further, we interrogated the proteome of three different wild-type P. aeruginosa strains, their eftM mutants, and these mutants complemented with eftM and conclude that trimethylation of EF-Tu by EftM does not impact EF-Tu's canonical function in translation. In addition to furthering our knowledge of this Pseudomonas virulence factor, this study provides an intriguing example of a protein with multiple layers of thermoregulation. eng Trimethylation of Elongation Factor-Tu by the Dual Thermoregulated Methyltransferase EftM Does Not Impact Its Canonical Function in Translation research article