Proteomic and phosphoproteomic profilings reveal distinct cellular responses during Tilapinevirus tilapiae entry and replication

Proteomic and phosphoproteomic profilings reveal distinct cellular responses during Tilapinevirus tilapiae entry and replication | 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 Proteomic and phosphoproteomic profilings reveal distinct cellular responses during Tilapinevirus tilapiae entry and replication Tuchakorn Lertwanakarn, Matepiya Khemthong, Piyathip Setthawong, and 4 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4730524/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Tilapia lake virus (TiLV) poses a significant threat to global tilapia aquaculture, as it causes high mortality rates and severe economic losses. Nevertheless, the molecular mechanisms of TiLV–host interactions remain poorly understood. We investigated the proteomic and phosphoproteomic changes in E-11 and RHTiB cell lines following TiLV infection. Using mass spectrometry-based analyses, we identified substantial alterations in protein expression and phosphorylation states, highlighted distinct cell line responses. In E-11 cells, TiLV infection suppressed the proteins involved in the Janus kinase–signal transducer and activators of transcription and Fas-associated death domain protein–tumor necrosis factor receptor associated factor pathways, leading to the activation of nucleotide oligomerization domain signaling and cellular apoptosis. The TiLV-infected RHTiB cells showed suppression of the host cellular metabolism through a reduction in protein phosphatase to facilitate early viral entry, while later stages of infection involved increased activity of myosin heavy chain 9 and enhanced host immune responses via the phosphorylation of ribosomal protein L17 and GTPase immunity-associated protein family member 7. These findings suggest that TiLV employs different strategies to manipulate host cellular pathways depending on the cell type. Further studies are essential to validate these findings and ultimately facilitate the development of effective antiviral strategies. Biological sciences/Microbiology/Virology Biological sciences/Biological techniques/Proteomic analysis Biological sciences/Cell biology/Cell signalling Full Text Additional Declarations No competing interests reported. Supplementary Files SupplementS1ProteomicE11RHTiB.xlsx SupplementS2PhosphoproteomicsE11RHTiB.xlsx SupplementS3ANOVAE11Proteomics.xlsx SupplementS4ANOVAE11Phosphoproteomics.xlsx SupplementS5AlteredproteinPhosphoproteinsE1110m24h.xlsx SupplementS6E11KeGG10m24h.xlsx SupplementS7ANOVARHTiBProteomics.xlsx SupplementS8ANOVARHTiBPhosphoproteomics.xlsx SupplementS9AlteredproteinPhosphoproteinsRHTiB10m24h.xlsx SupplementS10RHTiBKeGG10m24h.xlsx Graphicalabstract.tiff Cite Share Download PDF Status: Posted 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-4730524","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":335077612,"identity":"b0c3e1ba-e47f-40ce-8aa9-ea3a72cf804a","order_by":0,"name":"Tuchakorn Lertwanakarn","email":"","orcid":"","institution":"Kasetsart University","correspondingAuthor":false,"prefix":"","firstName":"Tuchakorn","middleName":"","lastName":"Lertwanakarn","suffix":""},{"id":335077613,"identity":"e89d3f87-371d-4525-b44c-13248283a00f","order_by":1,"name":"Matepiya Khemthong","email":"","orcid":"","institution":"Kasetsart University","correspondingAuthor":false,"prefix":"","firstName":"Matepiya","middleName":"","lastName":"Khemthong","suffix":""},{"id":335077614,"identity":"cce25e69-e1bf-49b9-85f6-17ddd6a6ddd9","order_by":2,"name":"Piyathip Setthawong","email":"","orcid":"","institution":"Kasetsart University","correspondingAuthor":false,"prefix":"","firstName":"Piyathip","middleName":"","lastName":"Setthawong","suffix":""},{"id":335077615,"identity":"57b3f356-258d-46f0-8f6d-1d3147f64278","order_by":3,"name":"Narumon Phaonakrop","email":"","orcid":"","institution":"National Center for Genetic Engineering and Biotechnology","correspondingAuthor":false,"prefix":"","firstName":"Narumon","middleName":"","lastName":"Phaonakrop","suffix":""},{"id":335077616,"identity":"f5840f0a-78a8-4584-b134-1af2874c03a2","order_by":4,"name":"Sittiruk Roytrakul","email":"","orcid":"","institution":"National Center for Genetic Engineering and Biotechnology","correspondingAuthor":false,"prefix":"","firstName":"Sittiruk","middleName":"","lastName":"Roytrakul","suffix":""},{"id":335077617,"identity":"780c47f5-247f-4ed2-82bc-4963a26e2cfe","order_by":5,"name":"Sekkarin Ploypetch","email":"","orcid":"","institution":"Mahidol University","correspondingAuthor":false,"prefix":"","firstName":"Sekkarin","middleName":"","lastName":"Ploypetch","suffix":""},{"id":335077618,"identity":"81da35a6-f87b-4888-8b36-0b58b68d4a92","order_by":6,"name":"Win Surachetpong","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAjElEQVRIiWNgGAWjYDACZhBRkQAkeEjScgaohY1oLSDA2EaKFoPjvAcYf85LS2yQ7z1ApJbDfAnMvNtyEhvY+BKI02J2mMeAmXFbBVALjwHxWhh/ziFVCwNvQw4JWuyBWg7zHEszbmPLIVKLZP8Zw4c/apJl+5nPEKkFBA6ACDbi1Y+CUTAKRsEoIAgA83YjvJ+CPqcAAAAASUVORK5CYII=","orcid":"","institution":"Kasetsart University","correspondingAuthor":true,"prefix":"","firstName":"Win","middleName":"","lastName":"Surachetpong","suffix":""}],"badges":[],"createdAt":"2024-07-12 13:11:29","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4730524/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4730524/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":64658732,"identity":"b6edd198-9acf-4d99-8372-680db8dfbde7","added_by":"auto","created_at":"2024-09-17 07:29:25","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1162432,"visible":true,"origin":"","legend":"","description":"","filename":"TiLVPhosphoproteomic.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4730524/v1_covered_7367da68-81e1-4857-bfc0-a22ffc2916e9.pdf"},{"id":61830548,"identity":"4c62131d-d992-4414-a5ea-cbc552cc6162","added_by":"auto","created_at":"2024-08-06 04:28:43","extension":"xlsx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":1323247,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementS1ProteomicE11RHTiB.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-4730524/v1/8b79136a46ff3442f7f1abfa.xlsx"},{"id":61830053,"identity":"5bd40318-2544-4106-b17f-b39983cb8e92","added_by":"auto","created_at":"2024-08-06 04:20:44","extension":"xlsx","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":1164706,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementS2PhosphoproteomicsE11RHTiB.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-4730524/v1/b81075c6dc2b3c8e2b6d55a7.xlsx"},{"id":61832011,"identity":"34f9568a-8b65-4ca6-ad48-1f8c5f229cf6","added_by":"auto","created_at":"2024-08-06 04:44:43","extension":"xlsx","order_by":3,"title":"","display":"","copyAsset":false,"role":"supplement","size":13210,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementS3ANOVAE11Proteomics.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-4730524/v1/6407e7e8a99635ae1d7c7e60.xlsx"},{"id":61831061,"identity":"7403a921-0991-4c07-82a9-d66f370d2099","added_by":"auto","created_at":"2024-08-06 04:36:43","extension":"xlsx","order_by":4,"title":"","display":"","copyAsset":false,"role":"supplement","size":17527,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementS4ANOVAE11Phosphoproteomics.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-4730524/v1/6ac7c288dff891fe82ba266a.xlsx"},{"id":61830045,"identity":"45730d87-cafc-486d-92af-aba2ae6413bf","added_by":"auto","created_at":"2024-08-06 04:20:43","extension":"xlsx","order_by":5,"title":"","display":"","copyAsset":false,"role":"supplement","size":18698,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementS5AlteredproteinPhosphoproteinsE1110m24h.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-4730524/v1/1dac87c9917fb2eef711d1dc.xlsx"},{"id":61830055,"identity":"fb30fd5f-65f0-4a7d-a3de-8d6d78ba42a3","added_by":"auto","created_at":"2024-08-06 04:20:44","extension":"xlsx","order_by":6,"title":"","display":"","copyAsset":false,"role":"supplement","size":22910,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementS6E11KeGG10m24h.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-4730524/v1/c338eb45387f66e9d50a886d.xlsx"},{"id":61830048,"identity":"3a4daf09-d064-4c88-a7ae-f0c793d6da8b","added_by":"auto","created_at":"2024-08-06 04:20:43","extension":"xlsx","order_by":7,"title":"","display":"","copyAsset":false,"role":"supplement","size":17226,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementS7ANOVARHTiBProteomics.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-4730524/v1/ae30ce27f4134e8e5afc8129.xlsx"},{"id":61830546,"identity":"b2ded3e2-06e6-48a7-9102-f1612ee83549","added_by":"auto","created_at":"2024-08-06 04:28:43","extension":"xlsx","order_by":8,"title":"","display":"","copyAsset":false,"role":"supplement","size":10449,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementS8ANOVARHTiBPhosphoproteomics.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-4730524/v1/60e167acf808c6f6986ba05e.xlsx"},{"id":61830051,"identity":"d77b4b4b-7682-49e6-a8d7-6c3282a8d04f","added_by":"auto","created_at":"2024-08-06 04:20:44","extension":"xlsx","order_by":9,"title":"","display":"","copyAsset":false,"role":"supplement","size":12513,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementS9AlteredproteinPhosphoproteinsRHTiB10m24h.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-4730524/v1/16a9eab42fc35f198a641515.xlsx"},{"id":61830052,"identity":"ec6c7179-63aa-4d53-bf2a-eab032356c0e","added_by":"auto","created_at":"2024-08-06 04:20:44","extension":"xlsx","order_by":10,"title":"","display":"","copyAsset":false,"role":"supplement","size":17429,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementS10RHTiBKeGG10m24h.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-4730524/v1/cb5a86de1252152fcef389b3.xlsx"},{"id":61830054,"identity":"e7cea72a-3774-49b5-be19-c3dbe9022419","added_by":"auto","created_at":"2024-08-06 04:20:44","extension":"tiff","order_by":11,"title":"","display":"","copyAsset":false,"role":"supplement","size":1090997,"visible":true,"origin":"","legend":"","description":"","filename":"Graphicalabstract.tiff","url":"https://assets-eu.researchsquare.com/files/rs-4730524/v1/55a0f23e89603937ae0d5c89.tiff"}],"financialInterests":"No competing interests reported.","formattedTitle":"Proteomic and phosphoproteomic profilings reveal distinct cellular responses during Tilapinevirus tilapiae entry and replication","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"","lastPublishedDoi":"10.21203/rs.3.rs-4730524/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4730524/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"Tilapia lake virus (TiLV) poses a significant threat to global tilapia aquaculture, as it causes high mortality rates and severe economic losses. Nevertheless, the molecular mechanisms of TiLV–host interactions remain poorly understood. We investigated the proteomic and phosphoproteomic changes in E-11 and RHTiB cell lines following TiLV infection. Using mass spectrometry-based analyses, we identified substantial alterations in protein expression and phosphorylation states, highlighted distinct cell line responses. In E-11 cells, TiLV infection suppressed the proteins involved in the Janus kinase–signal transducer and activators of transcription and Fas-associated death domain protein–tumor necrosis factor receptor associated factor pathways, leading to the activation of nucleotide oligomerization domain signaling and cellular apoptosis. The TiLV-infected RHTiB cells showed suppression of the host cellular metabolism through a reduction in protein phosphatase to facilitate early viral entry, while later stages of infection involved increased activity of myosin heavy chain 9 and enhanced host immune responses via the phosphorylation of ribosomal protein L17 and GTPase immunity-associated protein family member 7. These findings suggest that TiLV employs different strategies to manipulate host cellular pathways depending on the cell type. Further studies are essential to validate these findings and ultimately facilitate the development of effective antiviral strategies.","manuscriptTitle":"Proteomic and phosphoproteomic profilings reveal distinct cellular responses during Tilapinevirus tilapiae entry and replication","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-08-06 04:20:39","doi":"10.21203/rs.3.rs-4730524/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"b9b510ac-3ed3-40ec-b53d-b29bdfd15ddf","owner":[],"postedDate":"August 6th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":35486370,"name":"Biological sciences/Microbiology/Virology"},{"id":35486371,"name":"Biological sciences/Biological techniques/Proteomic analysis"},{"id":35486372,"name":"Biological sciences/Cell biology/Cell signalling"}],"tags":[],"updatedAt":"2024-09-17T07:21:16+00:00","versionOfRecord":[],"versionCreatedAt":"2024-08-06 04:20:39","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4730524","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4730524","identity":"rs-4730524","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}