The tRNA methylation activity and specificity of NSUN2 are independent of Archease
8 affiliations
- 1Center for Anatomy & Cell Biology, Medical University of Vienna, Vienna, Austria
- 2Department of Theoretical Chemistry, University of Vienna, Vienna, Austria
- 3Vienna Doctoral School in Chemistry (DoSChem), University of Vienna, Vienna, Austria
- 4Institute of Computer Science and Interdisciplinary Center for Bioinformatics, Leipzig University, Leipzig, Germany
- 5Institute of Molecular Biology, Biocenter, Medical University of Innsbruck, Innsbruck, Austria
- 6Max Perutz Labs, Medical University of Vienna, Vienna Biocenter Campus (VBC), Vienna, Austria
- 7Doctoral School, Vienna Biocenter PhD Program, a Doctoral School of the University of Vienna and the Medical University of Vienna, Vienna, Austria
- 8Faculty of Computer Science, Research Group Bioinformatics and Computational Biology, University of Vienna, Vienna, Austria
Abstract
Transfer RNAs (tRNAs) undergo extensive maturation, including modifications essential for translation efficiency and fidelity, as well as for structural stability. The modification cytosine-5 methylation (m5C) is catalyzed by NSUN2 at multiple positions within distinct structural regions of a subset of tRNAs, yet the determinants of its substrate specificity remain elusive. In archaea, the protein Archease has been implicated in modulating the cytosine specificity of the NSUN2 ortholog Trm4, whereas mammalian Archease was shown to be involved in pre-tRNA splicing. Here, we investigated whether mammalian Archease influences NSUN2 in a similar way like the archaeal enzyme. Using short hairpin RNA-mediated depletion of Archease in mouse embryonic stem cells and in vitro methylation assays with recombinant proteins, we assessed both the catalytic activity and substrate selectivity of NSUN2 in the presence or the absence of Archease. Our results indicate that neither the efficiency nor positional specificity of NSUN2-mediated tRNA m5C methylation is affected by Archease. These findings indicate that, unlike in archaea, Archease does not regulate tRNA methylation by NSUN2 in mammals, highlighting an evolutionary divergence in the functional role of Archease in tRNA maturation pathways.
Supplementary Information: The online version contains supplementary material available at https://doi.org/10.1038/s41598-026-63726-2.
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BibTeX
@article{stuchlik2026trna,
title = {{The tRNA methylation activity and specificity of NSUN2 are independent of Archease}},
author = {Valentina Stuchlik and Maria Waldl and Anming Huang and Moritz Kleinwächter and Anna Rocchegiani and Fatinah El-Isa and Ivo L. Hofacker and Javier Martinez and Alexandra Lusser and Elisa Vilardo},
journal = {Scientific Reports},
year = {2026},
month = jul,
doi = {10.1038/s41598-026-63726-2},
url = {https://doi.org/10.1038/s41598-026-63726-2},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - The tRNA methylation activity and specificity of NSUN2 are independent of Archease
AU - Valentina Stuchlik
AU - Maria Waldl
AU - Anming Huang
AU - Moritz Kleinwächter
AU - Anna Rocchegiani
AU - Fatinah El-Isa
AU - Ivo L. Hofacker
AU - Javier Martinez
AU - Alexandra Lusser
AU - Elisa Vilardo
PY - 2026
DA - 2026/07/29
JO - Scientific Reports
DO - 10.1038/s41598-026-63726-2
UR - https://doi.org/10.1038/s41598-026-63726-2
AB - Transfer RNAs (tRNAs) undergo extensive maturation, including modifications essential for translation efficiency and fidelity, as well as for structural stability. The modification cytosine-5 methylation (m5C) is catalyzed by NSUN2 at multiple positions within distinct structural regions of a subset of tRNAs, yet the determinants of its substrate specificity remain elusive. In archaea, the protein Archease has been implicated in modulating the cytosine specificity of the NSUN2 ortholog Trm4, whereas mammalian Archease was shown to be involved in pre-tRNA splicing. Here, we investigated whether mammalian Archease influences NSUN2 in a similar way like the archaeal enzyme. Using short hairpin RNA-mediated depletion of Archease in mouse embryonic stem cells and in vitro methylation assays with recombinant proteins, we assessed both the catalytic activity and substrate selectivity of NSUN2 in the presence or the absence of Archease. Our results indicate that neither the efficiency nor positional specificity of NSUN2-mediated tRNA m5C methylation is affected by Archease. These findings indicate that, unlike in archaea, Archease does not regulate tRNA methylation by NSUN2 in mammals, highlighting an evolutionary divergence in the functional role of Archease in tRNA maturation pathways. Supplementary Information: The online version contains supplementary material available at https://doi.org/10.1038/s41598-026-63726-2.
SN - 2045-2322
ER -