{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,1,16]],"date-time":"2026-01-16T01:51:46Z","timestamp":1768528306948,"version":"3.49.0"},"posted":{"date-parts":[[2025,10,13]]},"group-title":"Biochemistry","reference-count":57,"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\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,10,13]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>tRNA N\u2077-methylguanosine (m\u2077G46), installed by the METTL1\/WDR4 methyltransferase, stabilizes the tRNA pool supporting high proliferative rates. Accordingly, many cancer cell lines are vulnerable to METTL1 loss, underscoring its potential as a therapeutic target. We identify a subset of cell lines insensitive to METTL1 depletion and use oesophageal lines as a model to explore the mechanism. The METTL1-resistant adenocarcinoma line OE33 preserves tRNA balance upon m\u2077G46 deficiency by elevating dihydrouridylation at position 47 (D47). Depletion of DUS-L enzymes that catalyse D16\/D17\/D20\/D47 on tRNAs sensitises OE33 to METTL1 loss. Concomitant loss of dihydrouridylation and m\u2077G46 destabilizes the canonical L-shaped tRNA structure, disrupts coaxial helix stacking and anticodon-loop presentation, and reduces cellular fitness. Notably, METTL1-sensitive cell lines lack this compensatory pathway due to the deleterious effects of increasing their intrinsically low DUS3L levels. These findings reveal a DUS-L\u2013mediated buffering mechanism that supports tRNA homeostasis in METTL1-resistant cancers and highlight DUS3L abundance as a potential biomarker for predicting METTL1 sensitivity. Moreover, they suggest that combined targeting of DUS-L and METTL1 may provide a therapeutic strategy for overcoming resistance in cancers, exemplified in cell lines such as OE33.<\/jats:p>\n                <jats:sec>\n                  <jats:title>Highlights<\/jats:title>\n                  <jats:list list-type=\"bullet\">\n                    <jats:list-item>\n                      <jats:p>\n                        The oesophagus cancer line OE33 shows no proliferative defect upon METTL1\/WDR4 loss, despite complete lack of m\n                        <jats:sup>7<\/jats:sup>\n                        G46 on its tRNA.\n                      <\/jats:p>\n                    <\/jats:list-item>\n                    <jats:list-item>\n                      <jats:p>OE33 coordinates codon usage and abundance of cognate tRNAs in the absence of METTL1\/WDR4; the METTL1\/WDR4 sensitive line, OE21, fails to do so.<\/jats:p>\n                    <\/jats:list-item>\n                    <jats:list-item>\n                      <jats:p>Depleting dihydrouridine synthases (DUS-L) renders OE33 sensitive to METTL1\/WDR4 loss.<\/jats:p>\n                    <\/jats:list-item>\n                    <jats:list-item>\n                      <jats:p>OE33 responds to lack of METTL1\/WDR4 activity by upregulating DUS3L and its activity (D47) on tRNA; the sensitive line OE21 fails to do so.<\/jats:p>\n                    <\/jats:list-item>\n                    <jats:list-item>\n                      <jats:p>METTL1-sensitive lines have intrinsically low DUS3L and cannot sustain higher levels.<\/jats:p>\n                    <\/jats:list-item>\n                    <jats:list-item>\n                      <jats:p>DUS3L reveals as a potential biomarker for METTL1 sensitivity.<\/jats:p>\n                    <\/jats:list-item>\n                  <\/jats:list>\n                <\/jats:sec>","DOI":"10.1101\/2025.10.13.682082","type":"posted-content","created":{"date-parts":[[2025,10,14]],"date-time":"2025-10-14T03:05:12Z","timestamp":1760411112000},"source":"Crossref","is-referenced-by-count":0,"title":["Compensatory tRNA Modification by DUS3L Confers Resistance to METTL1 Loss in Oesophageal Cancer"],"prefix":"10.64898","author":[{"given":"Helena","family":"Santos-Rosa","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jonathan L.","family":"Price","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Georgia","family":"Tsagkogeorga","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alexandra","family":"Sapetschnig","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bianca","family":"Pierattini","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"David","family":"Jordan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Paula","family":"Milan-Rois","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Almudena","family":"Serrano-Benitez","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Katherine","family":"Stott","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Namshik","family":"Han","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tony","family":"Kouzarides","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Eric A.","family":"Miska","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2025101604250800000_2025.10.13.682082v1.1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkad1083"},{"key":"2025101604250800000_2025.10.13.682082v1.2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2017.05.045"},{"key":"2025101604250800000_2025.10.13.682082v1.3","first-page":"563","article-title":"Epitranscriptome\u2013epigenome interactions in development and disease mechanisms","volume":"41","year":"2025","journal-title":"Trends Genet"},{"key":"2025101604250800000_2025.10.13.682082v1.4","doi-asserted-by":"publisher","DOI":"10.3390\/genes15030374"},{"key":"2025101604250800000_2025.10.13.682082v1.5","doi-asserted-by":"publisher","DOI":"10.3390\/life6010003"},{"key":"2025101604250800000_2025.10.13.682082v1.6","doi-asserted-by":"publisher","DOI":"10.3390\/ijms19124080"},{"key":"2025101604250800000_2025.10.13.682082v1.7","doi-asserted-by":"publisher","DOI":"10.1017\/S1355838202024019"},{"key":"2025101604250800000_2025.10.13.682082v1.8","doi-asserted-by":"publisher","DOI":"10.1017\/S1355838200000364"},{"key":"2025101604250800000_2025.10.13.682082v1.9","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2005.10.036"},{"key":"2025101604250800000_2025.10.13.682082v1.10","doi-asserted-by":"publisher","DOI":"10.1261\/rna.052126.115"},{"key":"2025101604250800000_2025.10.13.682082v1.11","doi-asserted-by":"publisher","DOI":"10.1101\/gad.1654308"},{"key":"2025101604250800000_2025.10.13.682082v1.12","doi-asserted-by":"publisher","DOI":"10.1261\/rna.033654.112"},{"key":"2025101604250800000_2025.10.13.682082v1.13","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2018.06.001"},{"key":"2025101604250800000_2025.10.13.682082v1.14","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2021.06.031"},{"key":"2025101604250800000_2025.10.13.682082v1.15","doi-asserted-by":"publisher","DOI":"10.1186\/s13059-015-0779-x"},{"key":"2025101604250800000_2025.10.13.682082v1.16","first-page":"2508","article-title":"Mutations in WDR4 as a new cause of Galloway-Mowat syndrome","volume":"176","year":"2018","journal-title":"Am. 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