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Using tRNA sequencing, we profiled mature tRNAs during infections with human cytomegalovirus (HCMV) and SARS-CoV-2. HCMV-induced alterations in mature tRNA levels were predominantly virus-driven, with minimal influence from the cellular immune response. Certain post-transcriptional modifications, correlated with tRNA stability, were actively manipulated by HCMV. By contrast, SARS-CoV-2 caused minimal changes in mature tRNA levels or modifications. Comparing viral codon usage with proliferation- versus differentiation-associated codon-usage signatures in human genes revealed striking divergence. HCMV genes aligned with differentiation codon usage, whereas SARS-CoV-2 genes matched proliferation codon usage. Structural and gene-expression genes in both viruses showed strong adaptation to host tRNA pools. Finally, a systematic CRISPR screen of human tRNA genes and tRNA-modifying enzymes identified specific tRNAs and enzymes that either enhanced or restricted HCMV infectivity and influenced cellular growth. Together, these data define a dynamic interplay between the host tRNA landscape and viral infection, illuminating the mechanisms governing host\u2013virus interactions.<\/jats:p>","DOI":"10.1038\/s44320-025-00181-7","type":"journal-article","created":{"date-parts":[[2026,1,20]],"date-time":"2026-01-20T13:06:04Z","timestamp":1768914364000},"page":"519-536","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Essentiality and dynamic expression of the human tRNA pool during viral infection"],"prefix":"10.1038","volume":"22","author":[{"given":"Noa","family":"Aharon-Hefetz","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5442-0201","authenticated-orcid":false,"given":"Michal","family":"Schwartz","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Einav","family":"Aharon","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Noam","family":"Stern-Ginossar","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8096-5085","authenticated-orcid":false,"given":"Orna","family":"Dahan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3200-9344","authenticated-orcid":false,"given":"Yitzhak","family":"Pilpel","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2026,1,20]]},"reference":[{"key":"181_CR1","doi-asserted-by":"publisher","first-page":"1","DOI":"10.7554\/ELIFE.58461","volume":"9","author":"N Aharon-Hefetz","year":"2020","unstructured":"Aharon-Hefetz N, Frumkin I, Mayshar Y, Dahan O, Pilpel Y, Rak R (2020) Manipulation of the human tRNA pool reveals distinct tRNA sets that act in cellular proliferation or cell cycle arrest. 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