{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,15]],"date-time":"2026-10-15T19:40:47Z","timestamp":1792093247203,"version":"4.4.0"},"reference-count":51,"publisher":"Public Library of Science (PLoS)","issue":"7","license":[{"start":{"date-parts":[[2026,7,31]],"date-time":"2026-07-31T00:00:00Z","timestamp":1785456000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Key Research and Development Program of China","award":["2025YFC2608400"],"award-info":[{"award-number":["2025YFC2608400"]}]},{"DOI":"10.13039\/501100002858","name":"China Postdoctoral Science Foundation","doi-asserted-by":"publisher","award":["2022TQ0119"],"award-info":[{"award-number":["2022TQ0119"]}],"id":[{"id":"10.13039\/501100002858","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100013061","name":"Jilin Provincial Scientific and Technological Development Program","doi-asserted-by":"publisher","award":["20250601001RC"],"award-info":[{"award-number":["20250601001RC"]}],"id":[{"id":"10.13039\/501100013061","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plospathogens.org"],"crossmark-restriction":false},"short-container-title":["PLoS Pathog"],"abstract":"<jats:p>Getah virus (GETV), a mosquito-borne alphavirus, poses an emerging threat to public health with its increasingly broad host spectrum. While glycosaminoglycans (GAGs) serve as critical attachment factors for many alphaviruses and the low-density lipoprotein receptor (LDLR) facilitates the cellular entry of several members, the precise viral determinants governing these interactions and their implications for viral virulence remain poorly defined. Here, we introduced an H86Y substitution, a potential adaptive mutation site, within the E2 glycoprotein of GETV using reverse genetics. The H86Y mutant replicated more efficiently in mosquito C6\/36 cells but was consistently attenuated across several mammalian cell lines. In susceptible mouse models, H86Y infection led to reduced viral loads, milder histopathology, and lower inflammatory responses compared with the parental virus, yet still elicited robust protective immunity in adult mice. Mechanistically, a series of functional assays, including infection in GAG-deficient cells, decoy inhibition, co-immunoprecipitation, receptor overexpression and knockdown, and biolayer interferometry, demonstrated that the residue 86 in E2 glycoprotein is a critical determinant for GETV binding to both GAGs and LDLR. The H86Y mutation concurrently reduces these interactions, contributing the impairment of virus attachment and entry into mammalian cells. Furthermore, the GAG-binding site functionally overlaps with the LDLR interaction interface. In LDLR-deficient suckling mice, the impaired replication of H86Y persisted in examined tissues. However, pre-treatment with heparinase nearly completely eliminated this attenuation phenotype, further confirming that LDLR and GAG are the key host factors mediating attenuation phenotype for H86Y. In summary, the residue 86 of the GETV E2 glycoprotein represents a determinant of viral virulence, and an H86Y mutation attenuates GAGs and LDLR-dependent infection, providing mechanistic insights into alphavirus-host interactions and a potential target for antiviral and vaccine development.<\/jats:p>","DOI":"10.1371\/journal.ppat.1014453","type":"journal-article","created":{"date-parts":[[2026,7,31]],"date-time":"2026-07-31T17:33:51Z","timestamp":1785519231000},"page":"e1014453","update-policy":"https:\/\/doi.org\/10.1371\/journal.ppat.corrections_policy","source":"Crossref","is-referenced-by-count":1,"title":["The residue 86 of the Getah virus E2 glycoprotein mediates both glycosaminoglycan- and LDLR-dependent infection"],"prefix":"10.1371","volume":"22","author":[{"given":"Xiangshu","family":"Qiu","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rongguang","family":"Lu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiaxin","family":"Tian","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"He","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiyong","family":"Zhou","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mengsi","family":"Sun","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinyu","family":"Cao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiangyu","family":"Zhu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bocheng","family":"Liu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qihui","family":"Yu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuanyuan","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hualei","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ningyi","family":"Jin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4532-4466","authenticated-orcid":true,"given":"Huijun","family":"Lu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ning","family":"Shi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2026,7,31]]},"reference":[{"issue":"8","key":"ppat.1014453.ref001","doi-asserted-by":"crossref","first-page":"1167","DOI":"10.3390\/microorganisms8081167","article-title":"Epidemic alphaviruses: ecology, emergence and outbreaks","volume":"8","author":"SR Azar","year":"2020","journal-title":"Microorganisms"},{"issue":"4","key":"ppat.1014453.ref002","article-title":"Origin, genetic diversity, adaptive evolution and transmission dynamics of Getah virus","volume":"69","author":"N Shi","year":"2022","journal-title":"Transbound Emerg Dis"},{"issue":"1","key":"ppat.1014453.ref003","doi-asserted-by":"crossref","DOI":"10.1093\/ve\/veaf007","article-title":"Genomic surveillance and evolution of Getah virus","volume":"11","author":"J Shen","year":"2025","journal-title":"Virus Evol"},{"issue":"8","key":"ppat.1014453.ref004","doi-asserted-by":"crossref","first-page":"945","DOI":"10.3390\/pathogens11080945","article-title":"Getah virus (alphavirus): an emerging, spreading zoonotic virus","volume":"11","author":"B Li","year":"2022","journal-title":"Pathogens"},{"issue":"3","key":"ppat.1014453.ref005","doi-asserted-by":"crossref","first-page":"632","DOI":"10.1111\/tbed.12867","article-title":"An outbreak of Getah virus infection among pigs in China, 2017","volume":"65","author":"T Yang","year":"2018","journal-title":"Transbound Emerg Dis"},{"issue":"1","key":"ppat.1014453.ref006","doi-asserted-by":"crossref","first-page":"2548930","DOI":"10.1080\/21505594.2025.2548930","article-title":"The first discovery of the group IV GETV from Pangolin, China","volume":"16","author":"P Xiao","year":"2025","journal-title":"Virulence"},{"key":"ppat.1014453.ref007","doi-asserted-by":"crossref","first-page":"1583023","DOI":"10.3389\/fmicb.2025.1583023","article-title":"First report on identification and genetic characterization of Getah virus in wild boar in China","volume":"16","author":"J Huang","year":"2025","journal-title":"Front 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Song","year":"2023","journal-title":"Oncogene"},{"issue":"1","key":"ppat.1014453.ref029","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1186\/s12929-024-00995-x","article-title":"Attenuation of neurovirulence of chikungunya virus by a single amino acid mutation in viral E2 envelope protein","volume":"31","author":"H Chen","year":"2024","journal-title":"J Biomed Sci"},{"issue":"11","key":"ppat.1014453.ref030","doi-asserted-by":"crossref","DOI":"10.1371\/journal.ppat.1012700","article-title":"Attenuated Getah virus confers protection against multiple arthritogenic alphaviruses","volume":"20","author":"Z Jiang","year":"2024","journal-title":"PLoS Pathog"},{"issue":"5","key":"ppat.1014453.ref031","doi-asserted-by":"crossref","first-page":"2858","DOI":"10.1128\/JVI.03453-13","article-title":"Novel attenuated Chikungunya vaccine candidates elicit protective immunity in C57BL\/6 mice","volume":"88","author":"D Halleng\u00e4rd","year":"2014","journal-title":"J 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A"},{"issue":"7","key":"ppat.1014453.ref035","doi-asserted-by":"crossref","DOI":"10.1016\/j.cell.2019.04.006","article-title":"Cryo-EM structure of chikungunya virus in complex with the Mxra8 receptor","volume":"177","author":"K Basore","year":"2019","journal-title":"Cell"},{"issue":"2","key":"ppat.1014453.ref036","doi-asserted-by":"crossref","first-page":"85","DOI":"10.1016\/j.it.2023.11.006","article-title":"The many ways in which alphaviruses bind to cells","volume":"45","author":"S Raju","year":"2024","journal-title":"Trends Immunol"},{"issue":"10","key":"ppat.1014453.ref037","article-title":"Structure of Semliki Forest virus in complex with its receptor VLDLR","volume":"186","author":"D Cao","year":"2023","journal-title":"Cell"},{"issue":"4","key":"ppat.1014453.ref038","doi-asserted-by":"crossref","first-page":"250","DOI":"10.1016\/j.ibmb.2006.01.003","article-title":"Sequence analysis of the non-recurring C-terminal domains shows that insect lipoprotein receptors constitute a distinct group of LDL receptor family members","volume":"36","author":"KW Rodenburg","year":"2006","journal-title":"Insect Biochem Mol Biol"},{"issue":"11","key":"ppat.1014453.ref039","doi-asserted-by":"crossref","first-page":"1461","DOI":"10.1039\/D2AY01803A","article-title":"High sensitivity (zeptomole) detection of BODIPY-labelled heparan sulfate (HS) disaccharides by ion-paired RP-HPLC and LIF detection enables analysis of HS from mosquito midguts","volume":"15","author":"ML Maciej-Hulme","year":"2023","journal-title":"Anal Methods"},{"issue":"35","key":"ppat.1014453.ref040","doi-asserted-by":"crossref","first-page":"25376","DOI":"10.1074\/jbc.M704698200","article-title":"Mosquito heparan sulfate and its potential role in malaria infection and transmission","volume":"282","author":"P Sinnis","year":"2007","journal-title":"J Biol Chem"},{"issue":"2","key":"ppat.1014453.ref041","first-page":"001726","article-title":"Glycosaminoglycan binding by arboviruses: a cautionary tale","volume":"103","author":"MDH Alcorn","year":"2022","journal-title":"J Gen Virol"},{"issue":"758","key":"ppat.1014453.ref042","article-title":"A prM mutation that attenuates dengue virus replication in human cells enhances midgut infection in mosquitoes","volume":"16","author":"ANX Choi","year":"2024","journal-title":"Sci Transl Med"},{"issue":"5","key":"ppat.1014453.ref043","doi-asserted-by":"crossref","first-page":"744","DOI":"10.3390\/cells12050744","article-title":"Heparan Sulfates regulate axonal excitability and context generalization through Ca(2+)\/Calmodulin-dependent protein kinase II","volume":"12","author":"I Song","year":"2023","journal-title":"Cells"},{"issue":"10","key":"ppat.1014453.ref044","doi-asserted-by":"crossref","DOI":"10.1016\/j.celrep.2019.07.105","article-title":"Expression of the Mxra8 Receptor Promotes Alphavirus Infection and Pathogenesis in Mice and Drosophila","volume":"28","author":"R Zhang","year":"2019","journal-title":"Cell 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Miao","year":"2024","journal-title":"Vaccine"},{"issue":"13","key":"ppat.1014453.ref048","doi-asserted-by":"crossref","DOI":"10.1016\/j.heliyon.2024.e33432","article-title":"Infectivity and pathogenesis characterization of getah virus (GETV) strain via different inoculation routes in mice","volume":"10","author":"Z Jian","year":"2024","journal-title":"Heliyon"},{"issue":"10","key":"ppat.1014453.ref049","doi-asserted-by":"crossref","first-page":"2877","DOI":"10.1007\/s00705-018-3927-2","article-title":"Development of a TaqMan probe-based quantitative reverse transcription PCR assay for detection of Getah virus RNA","volume":"163","author":"N Shi","year":"2018","journal-title":"Arch Virol"},{"issue":"2","key":"ppat.1014453.ref050","doi-asserted-by":"crossref","first-page":"229","DOI":"10.1016\/j.virs.2022.02.004","article-title":"Molecular and serological surveillance of Getah virus in the Xinjiang Uygur Autonomous Region, China, 2017\u20132020","volume":"37","author":"N Shi","year":"2022","journal-title":"Virol Sin"},{"key":"ppat.1014453.ref051","article-title":"KVFinder-web: a web-based application for detecting and characterizing biomolecular cavities","volume":"51","author":"JVS Guerra","year":"2023","journal-title":"Nucleic Acids Res"}],"container-title":["PLOS 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