Loss of tRNA uridine thiolation affects mRNA translation, protein production, and sulfur-compound metabolism in Arabidopsis

Dannfald, A.; Cadoudal, A.; Carpentier, M.-C.; Merret, R.; Rigal, Q.; Vialaret, J.; Attina, A.; Kindermans, J.; Berrissou, C.; Koprivova, A.; David, A.; Hirtz, C.; Favory, J.-J.; Kopriva, S.; Drouard, L.; Deragon, J.-M.

doi:10.1101/2025.09.25.678457

bioRxiv · 10.1101/2025.09.25.678457

Loss of tRNA uridine thiolation affects mRNA translation, protein production, and sulfur-compound metabolism in Arabidopsis

Dannfald, A.·Cadoudal, A.·Carpentier, M.-C.·Merret, R.·Rigal, Q.·Vialaret, J.·Attina, A.·Kindermans, J.·Berrissou, C.·Koprivova, A.·David, A.·Hirtz, C.·Favory, J.-J.·Kopriva, S.·Drouard, L.·Deragon, J.-M.

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Abstract

The uridine at position 34 of tRNA anticodon loops is always modified at variable levels depending on environmental conditions, but a function for this highly conserved modification has not been firmly established. Using Arabidopsis thaliana, we show that the thiolation of U34 is a prerequisite for the subsequent modifications at position 32 and 37 of the tRNALys(UUU) anticodon loop, revealing a novel modification network. Surprisingly, the level of tRNALys(UUU) is strongly increased rather than reduced in the ctu1 or ctu2 mutant backgrounds that prevent these modifications. This suggests the existence of a regulatory feedback loop that drives the transcription of this specific tRNA gene family. Furthermore, we observed that the ability of the ribosome to decode AAA, GAA and CAA codons is impaired when the thiol group is lost, leading to a reduction in protein production, especially for genes enriched in these codons. Finally, we show that loss of tRNA thiolation results in variations in levels of many proteins involved in sulfur-compound metabolism and several sulfur-containing metabolites, suggesting that the level of tRNA thiolation may act as a sensor that regulates these processes.

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Dannfald, A., Cadoudal, A., Carpentier, M.-C., Merret, R., Rigal, Q., Vialaret, J., Attina, A., Kindermans, J., Berrissou, C., Koprivova, A., David, A., Hirtz, C., Favory, J.-J., Kopriva, S., Drouard, L., Deragon, J.-M.. 2025-09-25. Loss of tRNA uridine thiolation affects mRNA translation, protein production, and sulfur-compound metabolism in Arabidopsis. https://doi.org/10.1101/2025.09.25.678457

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