Preprint Article Version 1 This version is not peer-reviewed

Temperature-dependent tRNA Modifications in Bacillales

Version 1 : Received: 12 July 2024 / Approved: 12 July 2024 / Online: 12 July 2024 (14:51:40 CEST)

How to cite: Hoffmann, A.; Lorenz, C.; Fallmann, J.; Wolff, P.; Lechner, A.; Betat, H.; Mörl, M.; Stadler, P. F. Temperature-dependent tRNA Modifications in Bacillales. Preprints 2024, 2024071081. https://doi.org/10.20944/preprints202407.1081.v1 Hoffmann, A.; Lorenz, C.; Fallmann, J.; Wolff, P.; Lechner, A.; Betat, H.; Mörl, M.; Stadler, P. F. Temperature-dependent tRNA Modifications in Bacillales. Preprints 2024, 2024071081. https://doi.org/10.20944/preprints202407.1081.v1

Abstract

Transfer RNA (tRNA) modifications are essential for the temperature adaptation of thermophilic and psychrophilic organisms, as they control the rigidity and flexibility of transcripts. To further understand how specific tRNA modifications are adjusted to maintain functionality in response to temperature fluctuations, we investigated whether tRNA modifications represent an adaptation of bacteria to different growth temperatures (minimal, optimal, and maximal), focusing on closely related psychrophilic (P. halocryophilus and E. sibiricum), mesophilic (B. subtilis), and thermophilic (G. stearothermophilus) Bacillales. Utilizing an RNA sequencing approach combined with chemical pre-treatment of tRNA samples, we systematically profiled dihydrouridine (D), 4-thiouridine (s4U), 7-methyl-guanosine (m7G), and pseudouridine (Ψ) modifications at single-nucleotide resolution. Despite their close relation, each bacterium exhibited a unique tRNA modification profile. Our findings revealed increased tRNA modifications in the thermophilic bacteria at higher temperatures, particularly showing elevated levels of s4U8 and Ψ55 modifications compared to non-thermophilic bacteria, indicating a temperature-dependent regulation that may contribute to thermotolerance. Furthermore, we observed higher levels of D modifications in psychrophilic and mesophilic bacteria indicating an adaptive strategy for cold environments by enhancing local flexibility in tRNAs. Our method demonstrated high effectiveness in identifying tRNA modifications compared to an established tool, highlighting its potential for precise tRNA profiling studies.

Keywords

tRNA; RNA Modification; Bacteria; RNA sequencing; thermal adaption

Subject

Biology and Life Sciences, Biochemistry and Molecular Biology

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