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bioRxiv · 10.1101/2022.10.12.511899

An RNA modification enzyme directly senses reactive oxygen species for translational regulation in Enterococcus faecalis

Lee, W. L.·Sinha, A.·Lam, L. N.·Loo, H. L.·Liang, J.·Ho, P.·Liang, C.·Chan, C.·Begley, T.·Kline, K. A.·Dedon, P.
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Abstract

Bacteria possess elaborate systems to manage reactive oxygen and nitrogen species (ROS) arising from exposure to the mammalian immune system and environmental stresses. Here we report the discovery of an ROS-sensing RNA-modifying enzyme that regulates translation of stress-response proteins in the gut commensal and opportunistic pathogen Enterococcus faecalis. We analyzed the tRNA epitranscriptome of E. faecalis in response to reactive oxygen species (ROS) or sublethal doses of ROS-inducing antibiotics and identified large decreases in N2-methyladenosine (m2A) in both 23S ribosomal RNA and transfer RNA. This we determined to be due to ROS-mediated inactivation of the Fe-S cluster-containing methyltransferase, RlmN. Genetic knockout of RlmN gave rise to a proteome that mimicked the oxidative stress response, with increased levels of superoxide dismutase and decreased virulence proteins. While tRNA modifications are established to be dynamic for fine-tuning translation, here we report the first instance of a dynamically regulated, environmentally responsive rRNA modification. These studies lead to model in which RlmN serves as a redox-sensitive molecular switch, directly relaying oxidative stress to modulating translation through the rRNA and the tRNA epitranscriptome, revealing a new paradigm for understanding direct regulation of the proteome by RNA modifications.

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Lee, W. L., Sinha, A., Lam, L. N., Loo, H. L., Liang, J., Ho, P., Liang, C., Chan, C., Begley, T., Kline, K. A., Dedon, P.. 2022-10-13. An RNA modification enzyme directly senses reactive oxygen species for translational regulation in Enterococcus faecalis. https://doi.org/10.1101/2022.10.12.511899

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