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Trypanosoma brucei, the causative agent of human African trypanosomiasis, encodes a \u2018classical\u2019 La protein (TbLa) composed of a La-motif, two RNA recognition motifs (RRM1 and RRM2\u03b1), a C-terminal short basic motif (SBM), and a nuclear localization signal (NLS). In T. brucei, like in most eukaryotes, position 34 of tRNATyr, -Asp, -Asn and -His is modified with queuosine (Q34). The steady-state levels of queuosine-modified tRNA in the insect form (procyclic) of T. brucei can fluctuate dynamically depending on growth conditions, but the mechanism(s) controlling Q34 levels are not well understood. A well-established function of La is in precursor-tRNA 3\u2032-end metabolism, but in this work, we demonstrate that La also controls Q34-tRNA levels. Individual domain deletions showed that while deletion of La motif or RRM1 causes dysregulation of Q34-tRNA levels, no other domain plays a similar role. We also show that La is important for the normal balance of several additional tRNA modifications. These findings are discussed in the context of substrate competition between La and modification enzymes, also highlighting subcellular localization as a key determinant of tRNA function.<\/jats:p>","DOI":"10.1093\/nar\/gkaf594","type":"journal-article","created":{"date-parts":[[2025,7,10]],"date-time":"2025-07-10T10:49:28Z","timestamp":1752144568000},"source":"Crossref","is-referenced-by-count":5,"title":["Key RNA-binding domains in the La protein establish tRNA modification levels in <i>Trypanosoma brucei<\/i>"],"prefix":"10.1093","volume":"53","author":[{"ORCID":"https:\/\/orcid.org\/0009-0005-8639-7938","authenticated-orcid":false,"given":"Lankani","family":"Gunaratne","sequence":"first","affiliation":[{"name":"The Ohio State Biochemistry Program, The Ohio State University , Columbus, Ohio 43210 ,","place":["United States"]},{"name":"The Center for RNA Biology, The Ohio State University , Columbus, Ohio 43210 ,","place":["United States"]},{"name":"Cell Biology and Biochemistry and the Brown RNA Center Department of Molecular Biology, , 225 Dyer Street , Brown University, Providence, R\u00a0I\u00a0 02903 ,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Henry","family":"Moore","sequence":"additional","affiliation":[{"name":"University of California Department of Biomolecular Engineering, ,\u00a0 Santa Cruz , CA\u00a0 95064 ,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicholas","family":"Albaum","sequence":"additional","affiliation":[{"name":"Cell Biology and Biochemistry and the Brown RNA Center Department of Molecular Biology, , 225 Dyer Street , Brown University, Providence, R\u00a0I\u00a0 02903 ,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ananth","family":"Casius","sequence":"additional","affiliation":[{"name":"The Center for RNA Biology, The Ohio State University , Columbus, Ohio 43210 ,","place":["United States"]},{"name":"Cell Biology and Biochemistry and the Brown RNA Center Department of Molecular Biology, , 225 Dyer Street , Brown University, Providence, R\u00a0I\u00a0 02903 ,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jeremy","family":"Henderson","sequence":"additional","affiliation":[{"name":"New England Biolabs Inc , 240 County Road Ipswich , Massachusetts 01938 ,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alan","family":"Kessler","sequence":"additional","affiliation":[{"name":"Section on Molecular and Cell Biology, Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD), National Institutes of Health (NIH) , Bethesda , MD\u00a0 20892 ,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eva","family":"Heged\u0171sov\u00e1","sequence":"additional","affiliation":[{"name":"Institute of Parasitology, Biology Centre, Czech Academy of Sciences , \u010cesk\u00e9 Bud\u011bjovice,\u00a031 370 05,","place":["Czech Republic"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sneha","family":"Kulkarni","sequence":"additional","affiliation":[{"name":"Institute of Parasitology, Biology Centre, Czech Academy of Sciences , \u010cesk\u00e9 Bud\u011bjovice,\u00a031 370 05,","place":["Czech Republic"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Henry","family":"Arthur","sequence":"additional","affiliation":[{"name":"The Center for RNA Biology, The Ohio State University , Columbus, Ohio 43210 ,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8710-3675","authenticated-orcid":false,"given":"Robert\u00a0L","family":"Ross","sequence":"additional","affiliation":[{"name":"Thermo Fisher Scientific , Lexington , MA\u00a0 04241 ,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1019-7719","authenticated-orcid":false,"given":"Zden\u011bk","family":"Paris","sequence":"additional","affiliation":[{"name":"Institute of Parasitology, Biology Centre, Czech Academy of Sciences , \u010cesk\u00e9 Bud\u011bjovice,\u00a031 370 05,","place":["Czech Republic"]},{"name":"Faculty of Science, University of South Bohemia , \u010cesk\u00e9 Bud\u011bjovice,\u00a031 370 05,","place":["Czech Republic"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5209-0066","authenticated-orcid":false,"given":"Richard","family":"Maraia","sequence":"additional","affiliation":[{"name":"Section on Molecular and Cell Biology, Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD), National Institutes of Health (NIH) , Bethesda , MD\u00a0 20892 ,","place":["United 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