FTD-tau S320F mutation stabilizes local structure and allosterically promotes amyloid motif-dependent aggregation

FTD-tau S320F mutation stabilizes local structure and allosterically promotes amyloid motif-dependent aggregation | 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 FTD-tau S320F mutation stabilizes local structure and allosterically promotes amyloid motif-dependent aggregation Dailu Chen, Aleksandra Wosztyl, Vishruth Mullapudi, Sofia Bali, and 6 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-1950269/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 23 Mar, 2023 Read the published version in Nature Communications → Version 1 posted You are reading this latest preprint version Abstract Amyloid deposition of the microtubule-associated protein tau is a unifying theme in a multitude of neurodegenerative diseases. Disease-associated missense mutations in tau are associated with frontotemporal dementia (FTD) and enhance tau aggregation propensity. However, the molecular mechanism of how mutations in tau promote tau assembly into amyloids remains obscure. There is a need to understand how tau folds into pathogenic conformations to cause disease. Here we describe the structural mechanism for how an FTD-tau S320F mutation drives spontaneous aggregation. We use recombinant protein and synthetic peptide systems, computational modeling, cross-linking mass spectrometry, and cell models to investigate the mechanism of spontaneous aggregation of the S320F FTD-tau mutant. We discover that the S320F mutation drives the stabilization of a local hydrophobic cluster which allosterically exposes the 306 VQIVYK 311 amyloid motif. We identify a suppressor mutation that reverses the S320F aggregation phenotype through the reduction of S320F-based hydrophobic clustering in vitro and in cells. Finally, we use structure-based computational design to engineer rapidly aggregating tau sequences by optimizing nonpolar clusters in proximity to the S320 site revealing a new principle that governs the regulation of tau aggregation. We uncover a mechanism for regulating aggregation that balances transient nonpolar contacts within local protective structures or in longer-range interactions that sequester amyloid motifs. The introduction of a pathogenic mutation redistributes these transient interactions to drive spontaneous aggregation. We anticipate deeper knowledge of this process will permit control of tau aggregation into discrete structural polymorphs to aid design of reagents that can detect disease-specific tau conformations. Full Text Additional Declarations There is NO Competing Interest. Supplementary Files Sourcedata1.xlsx Source data 1 Sourcedata2.xlsx Source data 2 Sourcedata3.xlsx Source data 3 S320Fdraft20220810finalsupplementary.pdf NCOMMS2232292rs.pdf Reporting Summary Cite Share Download PDF Status: Published Journal Publication published 23 Mar, 2023 Read the published version in Nature Communications → 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-1950269","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":130206258,"identity":"409d4476-0cd4-47d0-a8dc-1f79a667389b","order_by":0,"name":"Dailu Chen","email":"","orcid":"","institution":"The University of Texas Southwestern Medical Center","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Dailu","middleName":"","lastName":"Chen","suffix":""},{"id":130206259,"identity":"9aada184-2fef-47d0-8b83-f43dfbf5eee9","order_by":1,"name":"Aleksandra Wosztyl","email":"","orcid":"https://orcid.org/0000-0003-1994-9040","institution":"The University of Texas Southwestern Medical Center","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Aleksandra","middleName":"","lastName":"Wosztyl","suffix":""},{"id":130206260,"identity":"53d42c7d-494b-4c23-83ba-809e14d9acc5","order_by":2,"name":"Vishruth Mullapudi","email":"","orcid":"https://orcid.org/0000-0002-5346-4242","institution":"The University of Texas Southwestern Medical Center","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Vishruth","middleName":"","lastName":"Mullapudi","suffix":""},{"id":130206261,"identity":"77ee6635-a352-4098-b541-9e4977a63552","order_by":3,"name":"Sofia Bali","email":"","orcid":"https://orcid.org/0000-0002-4046-7081","institution":"The University of Texas Southwestern Medical Center","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Sofia","middleName":"","lastName":"Bali","suffix":""},{"id":130206262,"identity":"b7aed910-83e6-4a58-bcd1-8ac7400a1e0f","order_by":4,"name":"Jaime Vacquer-Alicea","email":"","orcid":"","institution":"University of Texas Southwestern Medical Center","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Jaime","middleName":"","lastName":"Vacquer-Alicea","suffix":""},{"id":130206263,"identity":"664e3628-f73f-4c2d-b2ed-f4f8bd8974ec","order_by":5,"name":"Shamiram Melhem","email":"","orcid":"","institution":"Erasmus Medical Center","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Shamiram","middleName":"","lastName":"Melhem","suffix":""},{"id":130206264,"identity":"d2f34759-34b9-40f3-93d2-1c3cd10cea66","order_by":6,"name":"Harro Seelaar","email":"","orcid":"","institution":"Erasmus Medical Center","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Harro","middleName":"","lastName":"Seelaar","suffix":""},{"id":130206265,"identity":"2cdbf41f-2a58-49ec-8df2-de18ac4730e4","order_by":7,"name":"John van Swieten","email":"","orcid":"","institution":"Erasmus Medical Center","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"John","middleName":"van","lastName":"Swieten","suffix":""},{"id":130206266,"identity":"54fb534e-393a-4e83-a20e-d45478479f85","order_by":8,"name":"Marc Diamond","email":"","orcid":"https://orcid.org/0000-0002-8085-7770","institution":"The University of Texas Southwestern Medical Center","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Marc","middleName":"","lastName":"Diamond","suffix":""},{"id":130206267,"identity":"ab74a1a9-13da-423c-a0d5-9df545480a0d","order_by":9,"name":"Lukasz Joachimiak","email":"data:image/png;base64,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","orcid":"https://orcid.org/0000-0003-3061-5850","institution":"The University of Texas Southwestern Medical Center","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Lukasz","middleName":"","lastName":"Joachimiak","suffix":""}],"badges":[],"createdAt":"2022-08-10 18:55:49","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-1950269/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-1950269/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1038/s41467-023-37274-6","type":"published","date":"2023-03-23T04:00:00+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":25501902,"identity":"2222b0c9-0542-4eb7-918d-4285397e2686","added_by":"auto","created_at":"2022-08-22 16:03:39","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1609114,"visible":true,"origin":"","legend":"","description":"","filename":"S320Fdraft20220810finalmaintext.pdf","url":"https://assets-eu.researchsquare.com/files/rs-1950269/v1_covered.pdf"},{"id":25501897,"identity":"49b4a09e-99b9-49d4-81c8-6327fe2c91ae","added_by":"auto","created_at":"2022-08-22 16:03:31","extension":"xlsx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":539025,"visible":true,"origin":"","legend":"Source data 1","description":"","filename":"Sourcedata1.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-1950269/v1/10c8828f9f58bd094d810af0.xlsx"},{"id":25501898,"identity":"4b382f0d-8bc7-4726-88cb-671796451998","added_by":"auto","created_at":"2022-08-22 16:03:31","extension":"xlsx","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":19042,"visible":true,"origin":"","legend":"Source data 2","description":"","filename":"Sourcedata2.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-1950269/v1/e01102b35e9761dbae92fbc2.xlsx"},{"id":25501899,"identity":"abc6f0e7-078a-4edc-bae5-cc9924a36c48","added_by":"auto","created_at":"2022-08-22 16:03:32","extension":"xlsx","order_by":3,"title":"","display":"","copyAsset":false,"role":"supplement","size":617073,"visible":true,"origin":"","legend":"Source data 3","description":"","filename":"Sourcedata3.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-1950269/v1/3b4248d656224e0537905e1e.xlsx"},{"id":25501901,"identity":"b4d826c3-6501-4f3d-9615-57575899783b","added_by":"auto","created_at":"2022-08-22 16:03:32","extension":"pdf","order_by":4,"title":"","display":"","copyAsset":false,"role":"supplement","size":4509173,"visible":true,"origin":"","legend":"","description":"","filename":"S320Fdraft20220810finalsupplementary.pdf","url":"https://assets-eu.researchsquare.com/files/rs-1950269/v1/1329438fffda58933a96fa63.pdf"},{"id":25501900,"identity":"ae7a756e-171e-463d-b16e-18b38ecb455f","added_by":"auto","created_at":"2022-08-22 16:03:32","extension":"pdf","order_by":6,"title":"","display":"","copyAsset":false,"role":"supplement","size":2710939,"visible":true,"origin":"","legend":"Reporting Summary","description":"","filename":"NCOMMS2232292rs.pdf","url":"https://assets-eu.researchsquare.com/files/rs-1950269/v1/70c04b41766f7a7fba71621b.pdf"}],"financialInterests":"There is \u003cb\u003eNO\u003c/b\u003e Competing Interest.","formattedTitle":"FTD-tau S320F mutation stabilizes local structure and allosterically promotes amyloid motif-dependent aggregation","fulltext":[{"header":"Full Text","content":"This preprint is available for \u003ca href='/article/rs-1950269/latest.pdf' target='_blank'\u003edownload as a PDF\u003c/a\u003e."}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","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":"","lastPublishedDoi":"10.21203/rs.3.rs-1950269/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-1950269/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"Amyloid deposition of the microtubule-associated protein tau is a unifying theme in a multitude of neurodegenerative diseases. Disease-associated missense mutations in tau are associated with frontotemporal dementia (FTD) and enhance tau aggregation propensity. However, the molecular mechanism of how mutations in tau promote tau assembly into amyloids remains obscure. There is a need to understand how tau folds into pathogenic conformations to cause disease. Here we describe the structural mechanism for how an FTD-tau S320F mutation drives spontaneous aggregation. We use recombinant protein and synthetic peptide systems, computational modeling, cross-linking mass spectrometry, and cell models to investigate the mechanism of spontaneous aggregation of the S320F FTD-tau mutant. We discover that the S320F mutation drives the stabilization of a local hydrophobic cluster which allosterically exposes the \u003csup\u003e306\u003c/sup\u003eVQIVYK\u003csup\u003e311\u003c/sup\u003e amyloid motif. We identify a suppressor mutation that reverses the S320F aggregation phenotype through the reduction of S320F-based hydrophobic clustering in vitro and in cells. Finally, we use structure-based computational design to engineer rapidly aggregating tau sequences by optimizing nonpolar clusters in proximity to the S320 site revealing a new principle that governs the regulation of tau aggregation. We uncover a mechanism for regulating aggregation that balances transient nonpolar contacts within local protective structures or in longer-range interactions that sequester amyloid motifs. The introduction of a pathogenic mutation redistributes these transient interactions to drive spontaneous aggregation. We anticipate deeper knowledge of this process will permit control of tau aggregation into discrete structural polymorphs to aid design of reagents that can detect disease-specific tau conformations.","manuscriptTitle":"FTD-tau S320F mutation stabilizes local structure and allosterically promotes amyloid motif-dependent aggregation","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2022-08-22 16:03:30","doi":"10.21203/rs.3.rs-1950269/v1","editorialEvents":[],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"nature-communications","isNatureJournal":true,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"NCOMMS","sideBox":"Learn more about [Nature Communications](http://www.nature.com/ncomms/)","snPcode":"","submissionUrl":"https://mts-ncomms.nature.com/","title":"Nature Communications","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"ejp","reportingPortfolio":"Nature Communications","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"06cbb7a6-8130-4bf8-ac0d-1498c4aa76ef","owner":[],"postedDate":"August 22nd, 2022","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2023-03-24T07:08:06+00:00","versionOfRecord":{"articleIdentity":"rs-1950269","link":"https://doi.org/10.1038/s41467-023-37274-6","journal":{"identity":"nature-communications","isVorOnly":false,"title":"Nature Communications"},"publishedOn":"2023-03-23 04:00:00","publishedOnDateReadable":"March 23rd, 2023"},"versionCreatedAt":"2022-08-22 16:03:30","video":"","vorDoi":"10.1038/s41467-023-37274-6","vorDoiUrl":"https://doi.org/10.1038/s41467-023-37274-6","workflowStages":[]},"version":"v1","identity":"rs-1950269","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-1950269","identity":"rs-1950269","version":["v1"]},"buildId":"7rjqhiLT3MXkJMwkYKINL","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}