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Post-transcriptional modulation of the SigF regulon inMycobacterium smegmatisby the PhoH2 toxin-antitoxin

Abstract
PhoH2 proteins are highly conserved across bacteria and archaea yet their biological function is poorly characterised. We examined the growth profiles of Mycobacterium smegmatis strains mc²155 and mc²155 ΔphoH2 and observed the same growth profile and growth rate in a variety of conditions. In light of the comparable growth, we used RNAseq to provide a snapshot of the differences between the transcriptomes of M. smegmatis mc²155 and M. smegmatis mc²155 ΔphoH2 during normal growth. At 48 hours, elevated expression of the sigF regulon was observed in ΔphoH2 relative to wild type. In biochemical assays, PhoH2 showed activity toward sigF mRNA insinuating a role of PhoH2 in modulating the pool of sigF mRNA in the cell during normal growth, adding further complexity to the repertoire of reported mechanisms of post-translational regulation. Multiple copies of the preferred target site of PhoH2 were identified in loops of the sigF mRNA structure, leading us to propose a mechanism for the activity of PhoH2 that is initiated after assembly on specific single-stranded loops of RNA. We hypothesise that PhoH2 is a toxin-antitoxin that contributes to the regulation of SigF at a post-transcriptional level through targeted activity on sigF mRNA. This work presents the first evidence for post-transcriptional regulation of SigF along with the biological function of PhoH2 from M. smegmatis. This has implications for the highly conserved PhoH2 toxin-antitoxin module across the mycobacteria including the important human pathogen M. tuberculosis.
Type
Journal Article
Type of thesis
Series
Citation
Andrews, E. S. V., Rzoska-Smith, E., & Arcus, V. L. (2020). Post-transcriptional modulation of the SigF regulon inMycobacterium smegmatisby the PhoH2 toxin-antitoxin. PLOS ONE, 15(7). https://doi.org/10.1371/journal.pone.0236551
Date
2020
Publisher
Public Library Science
Degree
Supervisors
Rights
© 2020 Andrews et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.