{"paper_id":"391f6edf-e4c3-41f1-97e7-70a1530e875d","body_text":"Metabolic reprogramming of methylthioadenosine-dependent sulfur recycling is a major driver of CHIKV infection | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Metabolic reprogramming of methylthioadenosine-dependent sulfur recycling is a major driver of CHIKV infection Ursula Bilitewski, Jackson Muema, Myriam Friemelt, Heike Overwin-Moser, and 4 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8902794/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted You are reading this latest preprint version Abstract Viral replication is strongly dependent on host cell metabolites as primary sources of essential building blocks and due to the involvement of metabolites in the regulation of essential reactions. As chikungunya virus (CHIKV) is one of the emerging pathogens, we chose this virus to generate a metabolic profile of infected host cells. We detected that in particular 5ʹ-methylthioadenosine (MTA) concentrations were increased in infected cells. Additional studies showed that this increase was accompanied by increased methionine adenosyltransferase-2a (MAT2A) expression. In sulfur-depleted media, exogenously added MTA restored CHIKV replication more efficiently than its precursor S-adenosylmethionine (SAM), while inhibitions of MAT2A or of de novo cysteine (Cys) biosynthesis reduced viral infectivity. Moreover, infection with CHIKV upregulated the expression of the U34-tRNA methyltransferase ALKBH8 and deletion of ALKBH8 led to a reduction of virus replication, recapitulating Met-Cys deprivation effects. In line with the essential role of methyl donors (SAM) we proved the nanomolar potency of the S-adenosylhomocysteine hydrolase inhibitors DzNep and Adox for CHIKV replication. Thus, our results uncovered the relationship between metabolism and secondary regulatory effects for CHIKV replication. Biological sciences/Microbiology/Virology Biological sciences/Chemical biology Chikungunya virus pro-viral metabolites metabolic reprogramming host-pathogen interactions sulfur metabolism antiviral targets Full Text Additional Declarations There is NO Competing Interest. Supplementary Files SupplementaryfileS117022026.xlsx Supplementary File S1 SupplementaryfileS217022026.xlsx Supplementary file S2 SupplementaryfileS317022026.xlsx Supplementary file S3 SupplementaryfileS417022026.xlsx Supplementary file S4 SupplementaryTable1ubi.xlsx Supplementary Table 1 Cite Share Download PDF Status: Under Review Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {\"props\":{\"pageProps\":{\"initialData\":{\"identity\":\"rs-8902794\",\"acceptedTermsAndConditions\":true,\"allowDirectSubmit\":false,\"archivedVersions\":[],\"articleType\":\"Article\",\"associatedPublications\":[],\"authors\":[{\"id\":600240571,\"identity\":\"84f9f983-83dd-43c3-8088-38cf42b6f116\",\"order_by\":0,\"name\":\"Ursula Bilitewski\",\"email\":\"data:image/png;base64,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\",\"orcid\":\"\",\"institution\":\"Helmholtz-Centre for Infection Research\",\"correspondingAuthor\":true,\"prefix\":\"\",\"firstName\":\"Ursula\",\"middleName\":\"\",\"lastName\":\"Bilitewski\",\"suffix\":\"\"},{\"id\":600240572,\"identity\":\"e1c81d3c-126d-42f8-9619-7fab16f91a61\",\"order_by\":1,\"name\":\"Jackson Muema\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Helmholtz-Centre for Infection Research\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Jackson\",\"middleName\":\"\",\"lastName\":\"Muema\",\"suffix\":\"\"},{\"id\":600240573,\"identity\":\"f7ec99bf-5631-46f3-8af7-5885811da72d\",\"order_by\":2,\"name\":\"Myriam Friemelt\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Helmholtz-Centre for Infection Research\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Myriam\",\"middleName\":\"\",\"lastName\":\"Friemelt\",\"suffix\":\"\"},{\"id\":600240574,\"identity\":\"4aa2720f-54db-4aab-bed1-43508c2c99e9\",\"order_by\":3,\"name\":\"Heike Overwin-Moser\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Helmholtz-Centre for Infection Research\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Heike\",\"middleName\":\"\",\"lastName\":\"Overwin-Moser\",\"suffix\":\"\"},{\"id\":600240575,\"identity\":\"2a3d82ba-64e9-4de1-8360-421a1df28d48\",\"order_by\":4,\"name\":\"Ulrike Beutling\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Helmholtz-Centre for Infection Research\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Ulrike\",\"middleName\":\"\",\"lastName\":\"Beutling\",\"suffix\":\"\"},{\"id\":600240576,\"identity\":\"a9040b88-992f-46f8-aa3a-f19693a71a71\",\"order_by\":5,\"name\":\"Raimo Franke\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Helmholtz-Centre for Infection Research\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Raimo\",\"middleName\":\"\",\"lastName\":\"Franke\",\"suffix\":\"\"},{\"id\":600240577,\"identity\":\"2786c1be-36ad-4389-b495-675fc12b9674\",\"order_by\":6,\"name\":\"Peter Sandner\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Peter\",\"middleName\":\"\",\"lastName\":\"Sandner\",\"suffix\":\"\"},{\"id\":600240578,\"identity\":\"a9f56475-eaa3-4f65-b0b9-7a3bc457ec42\",\"order_by\":7,\"name\":\"Mark Broenstrup\",\"email\":\"\",\"orcid\":\"https://orcid.org/0000-0002-8971-7045\",\"institution\":\"Helmholtz Centre for Infection Research\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Mark\",\"middleName\":\"\",\"lastName\":\"Broenstrup\",\"suffix\":\"\"}],\"badges\":[],\"createdAt\":\"2026-02-17 16:16:55\",\"currentVersionCode\":1,\"declarations\":\"\",\"doi\":\"10.21203/rs.3.rs-8902794/v1\",\"doiUrl\":\"https://doi.org/10.21203/rs.3.rs-8902794/v1\",\"draftVersion\":[],\"editorialEvents\":[],\"editorialNote\":\"\",\"failedWorkflow\":false,\"files\":[{\"id\":104408414,\"identity\":\"1ee314aa-62a8-42f3-97a0-a584d7d9fa1f\",\"added_by\":\"auto\",\"created_at\":\"2026-03-11 12:42:22\",\"extension\":\"pdf\",\"order_by\":1,\"title\":\"\",\"display\":\"\",\"copyAsset\":false,\"role\":\"manuscript-pdf\",\"size\":2214512,\"visible\":true,\"origin\":\"\",\"legend\":\"Article File\",\"description\":\"\",\"filename\":\"Muemaetalmaintextandsupplfigtab.pdf\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-8902794/v1_covered_b0010b4b-9d91-48c7-83fd-2324ecc8fa1c.pdf\"},{\"id\":104040791,\"identity\":\"64dcc1b4-7d26-4237-93e2-0b0387608d42\",\"added_by\":\"auto\",\"created_at\":\"2026-03-06 04:33:23\",\"extension\":\"xlsx\",\"order_by\":0,\"title\":\"\",\"display\":\"\",\"copyAsset\":false,\"role\":\"supplement\",\"size\":85939,\"visible\":true,\"origin\":\"\",\"legend\":\"Supplementary File S1\",\"description\":\"\",\"filename\":\"SupplementaryfileS117022026.xlsx\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-8902794/v1/ff67b52022d123ebe935c113.xlsx\"},{\"id\":104040792,\"identity\":\"8052d5c4-081b-43e6-9023-2a3204f13953\",\"added_by\":\"auto\",\"created_at\":\"2026-03-06 04:33:23\",\"extension\":\"xlsx\",\"order_by\":1,\"title\":\"\",\"display\":\"\",\"copyAsset\":false,\"role\":\"supplement\",\"size\":52746,\"visible\":true,\"origin\":\"\",\"legend\":\"Supplementary file S2\",\"description\":\"\",\"filename\":\"SupplementaryfileS217022026.xlsx\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-8902794/v1/b109a4e2ccb6e56ee7e90aae.xlsx\"},{\"id\":104040795,\"identity\":\"c0e89dee-2bac-4694-af6a-734299e006aa\",\"added_by\":\"auto\",\"created_at\":\"2026-03-06 04:33:23\",\"extension\":\"xlsx\",\"order_by\":2,\"title\":\"\",\"display\":\"\",\"copyAsset\":false,\"role\":\"supplement\",\"size\":50742,\"visible\":true,\"origin\":\"\",\"legend\":\"Supplementary file S3\",\"description\":\"\",\"filename\":\"SupplementaryfileS317022026.xlsx\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-8902794/v1/44f49de4ee9fdef61e753b7f.xlsx\"},{\"id\":104402169,\"identity\":\"41e0d8e7-abc4-4c58-a1a6-07709cbd34d5\",\"added_by\":\"auto\",\"created_at\":\"2026-03-11 12:14:33\",\"extension\":\"xlsx\",\"order_by\":3,\"title\":\"\",\"display\":\"\",\"copyAsset\":false,\"role\":\"supplement\",\"size\":47461,\"visible\":true,\"origin\":\"\",\"legend\":\"Supplementary file S4\",\"description\":\"\",\"filename\":\"SupplementaryfileS417022026.xlsx\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-8902794/v1/c6d4bb08ed3b77c9c62bf541.xlsx\"},{\"id\":104402240,\"identity\":\"66681b73-bba1-4ef9-86b9-7d36face77ac\",\"added_by\":\"auto\",\"created_at\":\"2026-03-11 12:14:45\",\"extension\":\"xlsx\",\"order_by\":4,\"title\":\"\",\"display\":\"\",\"copyAsset\":false,\"role\":\"supplement\",\"size\":46058,\"visible\":true,\"origin\":\"\",\"legend\":\"Supplementary Table 1\",\"description\":\"\",\"filename\":\"SupplementaryTable1ubi.xlsx\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-8902794/v1/43586b78fe4ac958e9f6935c.xlsx\"}],\"financialInterests\":\"There is \\u003cb\\u003eNO\\u003c/b\\u003e Competing Interest.\",\"formattedTitle\":\"Metabolic reprogramming of methylthioadenosine-dependent sulfur recycling is a major driver of CHIKV infection\",\"fulltext\":[],\"fulltextSource\":\"\",\"fullText\":\"\",\"funders\":[],\"hasAdminPriorityOnWorkflow\":false,\"hasManuscriptDocX\":false,\"hasOptedInToPreprint\":true,\"hasPassedJournalQc\":\"\",\"hasAnyPriority\":false,\"hideJournal\":false,\"highlight\":\"\",\"institution\":\"\",\"isAcceptedByJournal\":false,\"isAuthorSuppliedPdf\":true,\"isDeskRejected\":\"\",\"isHiddenFromSearch\":false,\"isInQc\":false,\"isInWorkflow\":false,\"isPdf\":true,\"isPdfUpToDate\":true,\"isWithdrawnOrRetracted\":false,\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"identity\":\"nature-portfolio\",\"isNatureJournal\":true,\"hasQc\":false,\"allowDirectSubmit\":false,\"externalIdentity\":\"\",\"sideBox\":\"\",\"snPcode\":\"\",\"submissionUrl\":\"\",\"title\":\"Nature Portfolio\",\"twitterHandle\":\"\",\"acdcEnabled\":false,\"dfaEnabled\":false,\"editorialSystem\":\"ejp\",\"reportingPortfolio\":\"\",\"inReviewEnabled\":true,\"inReviewRevisionsEnabled\":false},\"keywords\":\"Chikungunya virus, pro-viral metabolites, metabolic reprogramming, host-pathogen interactions, sulfur metabolism, antiviral targets\",\"lastPublishedDoi\":\"10.21203/rs.3.rs-8902794/v1\",\"lastPublishedDoiUrl\":\"https://doi.org/10.21203/rs.3.rs-8902794/v1\",\"license\":{\"name\":\"CC BY 4.0\",\"url\":\"https://creativecommons.org/licenses/by/4.0/\"},\"manuscriptAbstract\":\"Viral replication is strongly dependent on host cell metabolites as primary sources of essential building blocks and due to the involvement of metabolites in the regulation of essential reactions. As chikungunya virus (CHIKV) is one of the emerging pathogens, we chose this virus to generate a metabolic profile of infected host cells. We detected that in particular 5ʹ-methylthioadenosine (MTA) concentrations were increased in infected cells. Additional studies showed that this increase was accompanied by increased methionine adenosyltransferase-2a (MAT2A) expression. In sulfur-depleted media, exogenously added MTA restored CHIKV replication more efficiently than its precursor S-adenosylmethionine (SAM), while inhibitions of MAT2A or of de novo cysteine (Cys) biosynthesis reduced viral infectivity. Moreover, infection with CHIKV upregulated the expression of the U34-tRNA methyltransferase ALKBH8 and deletion of ALKBH8 led to a reduction of virus replication, recapitulating Met-Cys deprivation effects. In line with the essential role of methyl donors (SAM) we proved the nanomolar potency of the S-adenosylhomocysteine hydrolase inhibitors DzNep and Adox for CHIKV replication. Thus, our results uncovered the relationship between metabolism and secondary regulatory effects for CHIKV replication.\",\"manuscriptTitle\":\"Metabolic reprogramming of methylthioadenosine-dependent sulfur recycling is a major driver of CHIKV infection\",\"msid\":\"\",\"msnumber\":\"\",\"nonDraftVersions\":[{\"code\":1,\"date\":\"2026-03-06 04:33:06\",\"doi\":\"10.21203/rs.3.rs-8902794/v1\",\"editorialEvents\":[],\"status\":\"published\",\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"identity\":\"communications-biology\",\"isNatureJournal\":true,\"hasQc\":false,\"allowDirectSubmit\":false,\"externalIdentity\":\"commsbio\",\"sideBox\":\"Learn more about [Communications Biology](http://www.nature.com/commsbio/)\",\"snPcode\":\"\",\"submissionUrl\":\"\",\"title\":\"Communications Biology\",\"twitterHandle\":\"\",\"acdcEnabled\":true,\"dfaEnabled\":true,\"editorialSystem\":\"ejp\",\"reportingPortfolio\":\"Communications Series\",\"inReviewEnabled\":true,\"inReviewRevisionsEnabled\":false}}],\"origin\":\"\",\"ownerIdentity\":\"fca4de30-6768-498f-8140-c5589f00db99\",\"owner\":[],\"postedDate\":\"March 6th, 2026\",\"published\":true,\"recentEditorialEvents\":[],\"rejectedJournal\":[],\"revision\":\"\",\"amendment\":\"\",\"status\":\"under-review\",\"subjectAreas\":[{\"id\":63873295,\"name\":\"Biological sciences/Microbiology/Virology\"},{\"id\":63873296,\"name\":\"Biological sciences/Chemical biology\"}],\"tags\":[],\"updatedAt\":\"2026-04-06T08:30:25+00:00\",\"versionOfRecord\":[],\"versionCreatedAt\":\"2026-03-06 04:33:06\",\"video\":\"\",\"vorDoi\":\"\",\"vorDoiUrl\":\"\",\"workflowStages\":[]},\"version\":\"v1\",\"identity\":\"rs-8902794\",\"journalConfig\":\"researchsquare\"},\"__N_SSP\":true},\"page\":\"/article/[identity]/[[...version]]\",\"query\":{\"redirect\":\"/article/rs-8902794\",\"identity\":\"rs-8902794\",\"version\":[\"v1\"]},\"buildId\":\"XKTyCvWXoU3ODBz1xrDgd\",\"isFallback\":false,\"isExperimentalCompile\":false,\"dynamicIds\":[84888],\"gssp\":true,\"scriptLoader\":[]}","source_license":"CC-BY-4.0","license_restricted":false}