Pseudouridines of tRNA Anticodon Stem-Loop Have Unexpected Role in Mutagenesis in Pseudomonas sp.

Abstract

Pseudouridines are known to be important for optimal translation. In this study we demonstrate an unexpected link between pseudouridylation of tRNA and mutation frequency in Pseudomonas species. We observed that the lack of pseudouridylation activity of pseudouridine synthases TruA or RluA elevates the mutation frequency in Pseudomonas putida 3 to 5-fold. The absence of TruA but not RluA elevates mutation frequency also in Pseudomonas aeruginosa. Based on the results of genetic studies and analysis of proteome data, the mutagenic effect of the pseudouridylation deficiency cannot be ascribed to the involvement of error-prone DNA polymerases or malfunctioning of DNA repair pathways. In addition, although the deficiency in TruA-dependent pseudouridylation made P. putida cells more sensitive to antimicrobial compounds that may cause oxidative stress and DNA damage, cultivation of bacteria in the presence of reactive oxygen species (ROS)-scavenging compounds did not eliminate the mutator phenotype. Thus, the elevated mutation frequency in the absence of tRNA pseudouridylation could be the result of a more specific response or, alternatively, of a cumulative effect of several small effects disturbing distinct cellular functions, which remain undetected when studied independently. This work suggests that pseudouridines link the translation machinery to mutation frequency.

Pseudouridines in RNAs: switching atoms means shifting paradigms

“…In detail, Ψ 38/39 appears in tandem with other modifications at U 34 , including mcm 5 U or s 2 U, which contributes to Glu/Gln‐rich protein translation. Moreover, the absence of Ψ 38/39 has an unexpected influence on the mutation frequency and wide spectrum of proteome changes in Pseudomonas spp [ 39 ]. Apart from these universally highly conserved Ψ‐sites, Ψs directly in the anticodon (positions 34, 35, and 36) are found only in certain tRNAs [ 40 ], such as Ψ 35 in tRNA Tyr , or Ψ 34/36 in tRNA Ile .…”