{
  "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.\n\nSupplementary Information: The online version contains supplementary material available at https://doi.org/10.1038/s41598-026-63726-2.",
  "abstract_shown": true,
  "affiliations": [
    "Center for Anatomy & Cell Biology, Medical University of Vienna, Vienna, Austria",
    "Department of Theoretical Chemistry, University of Vienna, Vienna, Austria",
    "Vienna Doctoral School in Chemistry (DoSChem), University of Vienna, Vienna, Austria",
    "Institute of Computer Science and Interdisciplinary Center for Bioinformatics, Leipzig University, Leipzig, Germany",
    "Institute of Molecular Biology, Biocenter, Medical University of Innsbruck, Innsbruck, Austria",
    "Max Perutz Labs, Medical University of Vienna, Vienna Biocenter Campus (VBC), Vienna, Austria",
    "Doctoral School, Vienna Biocenter PhD Program, a Doctoral School of the University of Vienna and the Medical University of Vienna, Vienna, Austria",
    "Faculty of Computer Science, Research Group Bioinformatics and Computational Biology, University of Vienna, Vienna, Austria"
  ],
  "article_type": "research article",
  "arxiv": null,
  "author_total": 10,
  "authors": [
    {
      "affs": [
        0
      ],
      "name": "Valentina Stuchlik"
    },
    {
      "affs": [
        0,
        1,
        2,
        3
      ],
      "name": "Maria Waldl"
    },
    {
      "affs": [
        4
      ],
      "name": "Anming Huang"
    },
    {
      "affs": [
        5,
        6
      ],
      "name": "Moritz Kleinwächter"
    },
    {
      "affs": [
        0
      ],
      "name": "Anna Rocchegiani"
    },
    {
      "affs": [
        0
      ],
      "name": "Fatinah El-Isa"
    },
    {
      "affs": [
        1,
        7
      ],
      "name": "Ivo L. Hofacker"
    },
    {
      "affs": [
        5
      ],
      "name": "Javier Martinez"
    },
    {
      "affs": [
        4
      ],
      "name": "Alexandra Lusser"
    },
    {
      "affs": [
        0
      ],
      "name": "Elisa Vilardo",
      "orcid": "0000-0003-2733-6009"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/4e2c8jj8c8ysy.bib",
  "cited_by": 0,
  "doi": "10.1038/s41598-026-63726-2",
  "fulltext_url": "https://pmc.ncbi.nlm.nih.gov/articles/PMC13713551/",
  "id": "4e2c8jj8c8ysy",
  "issns": [
    "2045-2322"
  ],
  "landing_url": "https://doi.org/10.1038/s41598-026-63726-2",
  "language": "en",
  "licence": "CC-BY-4.0",
  "licence_label": "CC BY 4.0",
  "licence_source": "article",
  "licence_url": "https://creativecommons.org/licenses/by/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "heading",
      "Article"
    ],
    [
      "openalex",
      "1312|RNA modifications and cancer"
    ]
  ],
  "openalex": "W7171666455",
  "pdf_url": "https://www.nature.com/articles/s41598-026-63726-2_reference.pdf",
  "pmcid": "PMC13713551",
  "pmid": null,
  "published": "2026-07-29",
  "publisher": null,
  "ris": "https://research.draconitic.com/paper/4e2c8jj8c8ysy.ris",
  "sources": [
    {
      "first_seen": "2026-10-31T12:24:05Z",
      "native_id": "PMC13713551",
      "source": "pmc",
      "url": "https://doi.org/10.1038/s41598-026-63726-2"
    }
  ],
  "subjects": [
    {
      "field": "biochemistry",
      "origin": "openalex:1312|RNA modifications and cancer",
      "subfield": "molecular-biology"
    }
  ],
  "title": "The tRNA methylation activity and specificity of NSUN2 are independent of Archease",
  "title_text": "The tRNA methylation activity and specificity of NSUN2 are independent of Archease",
  "type": "peer-reviewed",
  "updated_at": "2026-11-01T12:21:12Z",
  "url": "https://research.draconitic.com/paper/4e2c8jj8c8ysy",
  "venue": "Scientific Reports",
  "versions": []
}