{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T12:46:23Z","timestamp":1785415583339,"version":"3.56.0"},"posted":{"date-parts":[[2025,10,13]]},"group-title":"Microbiology","reference-count":141,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,10,13]],"date-time":"2025-10-13T00:00:00Z","timestamp":1760313600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"award":["R35GM156215"],"award-info":[{"award-number":["R35GM156215"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]},{"award":["R01GM70641"],"award-info":[{"award-number":["R01GM70641"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]},{"award":["R01GM110588"],"award-info":[{"award-number":["R01GM110588"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]},{"award":["R01ES026856"],"award-info":[{"award-number":["R01ES026856"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]},{"award":["R35GM131767"],"award-info":[{"award-number":["R35GM131767"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]},{"id":[{"id":"https:\/\/ror.org\/02ebx7v45","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,11,8]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  tRNA modifications are central to bacterial translational control. Here, we integrated genetics, mass spectrometry, epitranscriptomics, and comparative genomics to map the tRNA modification genes of the Gram-positive pathogens\n                  <jats:italic>Streptococcus mutans<\/jats:italic>\n                  and\n                  <jats:italic>Streptococcus pneumoniae<\/jats:italic>\n                  . Both species show a marked loss of modifications dependent on Fe\u2013S enzymes, consistent with a broader trend of Fe\u2013S enzyme reduction in Streptococcus central metabolism. In addition, the D, m\n                  <jats:sup>1<\/jats:sup>\n                  A, m\n                  <jats:sup>7<\/jats:sup>\n                  G, t\n                  <jats:sup>6<\/jats:sup>\n                  A, and i\n                  <jats:sup>6<\/jats:sup>\n                  A modifications were mapped in\n                  <jats:italic>S. pneumoniae<\/jats:italic>\n                  tRNAs, and we confirmed that a unique DusB1 enzyme is responsible for the insertion of all the detectable D modifications. We uncovered differences in queuosine (Q) metabolism: while\n                  <jats:italic>S. mutans<\/jats:italic>\n                  synthesizes Q de novo,\n                  <jats:italic>S. pneumoniae<\/jats:italic>\n                  instead salvages preQ\u2081 and accumulates the epoxy-Q precursor, a strategy shared with multiple other Streptococci as revealed by analysis of Q pathways in 1,599 sequenced streptococcal genomes. Comparative essentiality profiling of modification genes revealed notable differences, including the essentiality of the NLJ-threonylcarbamoyladenosine (tLJA) synthesis enzyme TsaE in\n                  <jats:italic>S. pneumoniae<\/jats:italic>\n                  but not in\n                  <jats:italic>S. mutans<\/jats:italic>\n                  , which was confirmed by genetic studies. We found that suppressor mutations in\n                  <jats:italic>asnS<\/jats:italic>\n                  encoding asparaginyl-tRNA synthetase (AsnRS) restored viability to \u0394\n                  <jats:italic>tsaE<\/jats:italic>\n                  mutants, albeit with reduced growth. Our finding highlights the functional importance of modifications in the recognition of tRNAs by aminoacyl-tRNA synthetases.\n                <\/jats:p>","DOI":"10.1101\/2025.10.13.681863","type":"posted-content","created":{"date-parts":[[2025,10,14]],"date-time":"2025-10-14T03:20:15Z","timestamp":1760412015000},"source":"Crossref","is-referenced-by-count":1,"title":["tRNA Modification Landscapes in Streptococci: Shared Losses and Clade-Specific Adaptations"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0849-874X","authenticated-orcid":false,"given":"Ho-Ching 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