{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,8,7]],"date-time":"2026-08-07T16:51:32Z","timestamp":1786121492021,"version":"build-2736575974"},"posted":{"date-parts":[[2026,7,28]]},"group-title":"Molecular Biology","reference-count":35,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,7,28]],"date-time":"2026-07-28T00:00:00Z","timestamp":1785196800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000057","name":"National Institute of General Medical Sciences","doi-asserted-by":"crossref","award":["R35152086"],"award-info":[{"award-number":["R35152086"]}],"id":[{"id":"10.13039\/100000057","id-type":"DOI","asserted-by":"crossref"}]},{"name":"Office of Research Infrastructure Programs","award":["S10OD032364"],"award-info":[{"award-number":["S10OD032364"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,8,4]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>Like other enveloped RNA viruses, alphaviruses synthesize and assemble their envelope glycoproteins at the endoplasmic reticulum (ER) membrane. There, protein-conducting channels called translocons provide nascent proteins access to the membrane and allow them to fold into their correct shapes. We previously showed that a hydrophobic segment in the Sindbis virus structural polyprotein forms cotranslational interactions with the translocon that enhance \u22121 programmed ribosomal frameshifting (\u22121PRF), a recoding event that regulates polyprotein biogenesis. Recent discoveries concerning translocon remodeling suggest this segment, which corresponds to the second transmembrane domain of the E2 protein, could serve as a signal that recruits the multipass translocon (MPT). Here, we show that knocking out certain components of the MPT increases \u22121PRF efficiency. These differences in recoding coincide with changes in the membrane topology of the nascent polyprotein and in its downstream proteolytic processing in a manner that ultimately reduces viral fitness. Together, our results indicate that \u22121PRF and spike protein maturation in alphaviruses is tuned by the dynamic remodeling of the ER translocon. Such coupling could allow polyprotein biogenesis to adapt to different stages of viral replication and to distinct host or vector environments.<\/jats:p>","DOI":"10.64898\/2026.07.27.741003","type":"posted-content","created":{"date-parts":[[2026,7,28]],"date-time":"2026-07-28T10:00:15Z","timestamp":1785232815000},"source":"Crossref","is-referenced-by-count":0,"title":["Translocon Remodeling Modulates Ribosomal Frameshifting and the Maturation of the Sindbis Virus Structural Polyprotein"],"prefix":"10.64898","author":[{"given":"Antonio","family":"Bonifasi","sequence":"first","affiliation":[{"name":"The James Tarpo Jr. and Margaret Tarpo Department of Chemistry, Purdue University, West Lafayette, Indiana, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gayani Apsara","family":"Ranasinghe","sequence":"additional","affiliation":[{"name":"Department of Molecular and Cellular Biology, Indiana University, Bloomington, Indiana, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ashish R.","family":"Jhangiani","sequence":"additional","affiliation":[{"name":"The James Tarpo Jr. and Margaret Tarpo Department of Chemistry, Purdue University, West Lafayette, Indiana, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Venkatesh P","family":"Thirumalaikumar","sequence":"additional","affiliation":[{"name":"Bindley Bioscience Institute, Purdue University, West Lafayette, IN USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brianna","family":"Corman","sequence":"additional","affiliation":[{"name":"Weldon School of Biomedical Engineering, Purdue University, West Lafayette, IN, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J. Paul","family":"Robinson","sequence":"additional","affiliation":[{"name":"Weldon School of Biomedical Engineering, Purdue University, West Lafayette, IN, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Suchetana","family":"Mukhopadhyay","sequence":"additional","affiliation":[{"name":"Department of Biology, Indiana University, Bloomington, Indiana, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0955-7633","authenticated-orcid":false,"given":"Jonathan P.","family":"Schlebach","sequence":"additional","affiliation":[{"name":"The James Tarpo Jr. and Margaret Tarpo Department of Chemistry, Purdue University, West Lafayette, Indiana, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026080621502069000_2026.07.27.741003v2.1","doi-asserted-by":"crossref","first-page":"585","DOI":"10.1038\/s41579-025-01177-8","article-title":"Chikungunya virus and other emerging arthritogenic alphaviruses","volume":"23","year":"2025","journal-title":"Nat. Rev. Microbiol"},{"key":"2026080621502069000_2026.07.27.741003v2.2","doi-asserted-by":"publisher","DOI":"10.1038\/s41579-022-00825-7"},{"key":"2026080621502069000_2026.07.27.741003v2.3","doi-asserted-by":"crossref","first-page":"30","DOI":"10.3390\/v18010030","article-title":"Hidden in Plain Sight: Alphavirus Persistence and Its Potential for Driving Chronic Pathogenesis","volume":"18","year":"2025","journal-title":"Viruses"},{"key":"2026080621502069000_2026.07.27.741003v2.4","doi-asserted-by":"crossref","first-page":"299","DOI":"10.1146\/annurev-virology-093022-010359","article-title":"A Structural Perspective on the Alphavirus Life Cycle","volume":"12","year":"2025","journal-title":"Annu. Rev. Virol"},{"key":"2026080621502069000_2026.07.27.741003v2.5","doi-asserted-by":"crossref","first-page":"382","DOI":"10.3390\/v15020382","article-title":"Sequelae and Animal Modeling of Encephalitic Alphavirus Infections","volume":"15","year":"2023","journal-title":"Viruses"},{"key":"2026080621502069000_2026.07.27.741003v2.6","doi-asserted-by":"crossref","first-page":"1301","DOI":"10.1128\/jvi.61.5.1301-1309.1987","article-title":"Processing of the Semliki Forest Virus Structural Polyprotein: Role of the Capsid Protease","volume":"61","year":"1987","journal-title":"J. Virol"},{"key":"2026080621502069000_2026.07.27.741003v2.7","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.64.9.4346-4355.1990"},{"key":"2026080621502069000_2026.07.27.741003v2.8","doi-asserted-by":"crossref","first-page":"499","DOI":"10.1111\/j.1432-1033.1984.tb08130.x","article-title":"Role of signal recognition particle in the membrane assembly of Sindbis viral glycoproteins","volume":"140","year":"1984","journal-title":"Eur. J. Biochem"},{"key":"2026080621502069000_2026.07.27.741003v2.9","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.77.5.2981-2989.2003"},{"key":"2026080621502069000_2026.07.27.741003v2.10","doi-asserted-by":"publisher","DOI":"10.1006\/viro.1996.0229"},{"key":"2026080621502069000_2026.07.27.741003v2.11","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.85.21.8052"},{"key":"2026080621502069000_2026.07.27.741003v2.12","first-page":"37","article-title":"Regulators of Viral Frameshifting: More Than RNA Influences Translation Events","volume":"31","year":"2026","journal-title":"Annu. Rev. Virol"},{"key":"2026080621502069000_2026.07.27.741003v2.13","doi-asserted-by":"publisher","DOI":"10.1186\/1743-422X-5-108"},{"key":"2026080621502069000_2026.07.27.741003v2.14","doi-asserted-by":"crossref","first-page":"228","DOI":"10.3390\/v9080228","article-title":"Disentangling the frames, the state of research on the alphavirus 6K and TF proteins","volume":"9","year":"2017","journal-title":"Viruses"},{"key":"2026080621502069000_2026.07.27.741003v2.15","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2010.01.044"},{"key":"2026080621502069000_2026.07.27.741003v2.16","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.00449-13"},{"key":"2026080621502069000_2026.07.27.741003v2.17","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.02000-16"},{"key":"2026080621502069000_2026.07.27.741003v2.18","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkw530"},{"key":"2026080621502069000_2026.07.27.741003v2.19","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.RA120.012706"},{"key":"2026080621502069000_2026.07.27.741003v2.20","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1713166114"},{"key":"2026080621502069000_2026.07.27.741003v2.21","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkab1172"},{"key":"2026080621502069000_2026.07.27.741003v2.22","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-022-05336-2"},{"key":"2026080621502069000_2026.07.27.741003v2.23","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-022-05330-8"},{"key":"2026080621502069000_2026.07.27.741003v2.24","doi-asserted-by":"crossref","first-page":"2517","DOI":"10.1038\/s41594-025-01691-6","article-title":"Global analysis of translocon remodeling during protein synthesis at the ER","volume":"32","year":"2025","journal-title":"Nat. Struct. Mol. Biol"},{"key":"2026080621502069000_2026.07.27.741003v2.25","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-022-05638-5"},{"key":"2026080621502069000_2026.07.27.741003v2.26","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.52760"},{"key":"2026080621502069000_2026.07.27.741003v2.27","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.biophys.37.032807.125904"},{"key":"2026080621502069000_2026.07.27.741003v2.28","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2014.04.041"},{"key":"2026080621502069000_2026.07.27.741003v2.29","doi-asserted-by":"publisher","DOI":"10.1038\/nature13428"},{"key":"2026080621502069000_2026.07.27.741003v2.30","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2015.02.003"},{"key":"2026080621502069000_2026.07.27.741003v2.31","doi-asserted-by":"publisher","DOI":"10.1016\/j.virol.2020.01.001"},{"key":"2026080621502069000_2026.07.27.741003v2.32","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-020-2624-y"},{"key":"2026080621502069000_2026.07.27.741003v2.33","doi-asserted-by":"publisher","DOI":"10.1021\/acssynbio.3c00355"},{"key":"2026080621502069000_2026.07.27.741003v2.34","doi-asserted-by":"crossref","unstructured":"Carmody, P. J. et al. Ribosomal frameshifting selectively modulates the assembly, function, and pharmacological rescue of a misfolded CFTR variant. Proc. Natl. Acad. Sci. U. S. A. 121, (2024).","DOI":"10.1073\/pnas.2414768121"},{"key":"2026080621502069000_2026.07.27.741003v2.35","doi-asserted-by":"crossref","unstructured":"Carmody, P. J. et al. Feedback from the Nascent Chain Triggers Ribosomal Frameshifting and Transcript Decay. bioRxiv (2025).","DOI":"10.64898\/2025.12.23.696290"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.64898\/2026.07.27.741003","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,7]],"date-time":"2026-08-07T04:50:46Z","timestamp":1786078246000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.64898\/2026.07.27.741003"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,28]]},"references-count":35,"URL":"https:\/\/doi.org\/10.64898\/2026.07.27.741003","relation":{},"subject":[],"published":{"date-parts":[[2026,7,28]]},"subtype":"preprint"}}