Identification of residues in Lassa virus glycoprotein 1 involved in receptor switch

Jiao Guo; Yi Wan; Yang Liu; Xiaoying Jia; Siqi Dong; Gengfu Xiao; Wei Wang

doi:10.1016/j.virs.2024.06.001

August 2024

Jiao Guo, Yi Wan, Yang Liu, Xiaoying Jia, Siqi Dong, Gengfu Xiao and Wei Wang. Identification of residues in Lassa virus glycoprotein 1 involved in receptor switch[J]. Virologica Sinica, 2024, 39(4): 600-608. doi: 10.1016/j.virs.2024.06.001

Citation: Jiao Guo, Yi Wan, Yang Liu, Xiaoying Jia, Siqi Dong, Gengfu Xiao, Wei Wang. Identification of residues in Lassa virus glycoprotein 1 involved in receptor switch .VIROLOGICA SINICA, 2024, 39(4) : 600-608. http://dx.doi.org/10.1016/j.virs.2024.06.001

拉沙病毒包膜糖蛋白GP1亚基参与受体切换关键位点的鉴定

郭娇 ^a,c ,
万仪 ^a,b ,
刘洋 ^a ,
贾潇瑛 ^a,b ,
董思琦 ^a,b ,
肖庚富 ^a,b ,
王薇 ^a,b,,

cstr: 32224.14.j.virs.2024.06.001

a. 中国科学院武汉病毒研究所;
b. 中国科学院大学;
c. 西安医学院

通讯作者： 王薇, wangwei@wh.iov.cn
收稿日期： 2024-01-18
录用日期： 2024-05-31

摘要

拉沙病毒（LASV）导致拉沙出血热，LASV感染细胞时从第一受体 α-DG切换至第二受体 LAMP1，而与其亲缘关系高的沙粒病毒模式病毒淋巴细胞性脉络丛脑膜炎病毒 (LCMV) 不发生受体切换。据报道，LASV-GP1中，在沙粒病毒科保守的组氨酸三联体参与受体切换。为了验证其它非保守残基是否也参与受体切换，我们基于序列和构象比对，构建了一系列 LASV-GP1 突变体蛋白，并测试它们与LAMP1的结合。结果发现，其中四个位点（L84、K88、L107 和 H170）在受体切换中发挥重要作用。将上述四个氨基酸位点分别突变为 LCMV-GP1上的相应氨基酸，均会导致LASV-GP1与LAMP1的结合力降低。此外，所有突变都会降低GPC在pH 4.5 和pH 5.2时的膜融合效率；其中，L84N 和 K88E还显著降低LASV假型病毒感染效率。通过对于LAMP1结合力的考察，发现四种突变体与 LAMP1的亲和力均下降，顺序为 L84N > L107F > K88E > H170S。本研究首次报道的这四个氨基酸位点对深入研究LASV受体转换和免疫逃逸的发生机制、开发抗LASV感染药物具有重要的参考意义。
- 拉沙病毒
- , LAMP1
- , 糖蛋白
- , 受体切换
- , 膜融合

Identification of residues in Lassa virus glycoprotein 1 involved in receptor switch

Jiao Guo ^a,c ,
Yi Wan ^a,b ,
Yang Liu ^a ,
Xiaoying Jia ^a,b ,
Siqi Dong ^a,b ,
Gengfu Xiao ^a,b ,
Wei Wang ^a,b,,

a. State Key Laboratory of Virology, Wuhan Institute of Virology, Center for Biosafety Mega-Science, Chinese Academy of Sciences, Wuhan 430071, China;
b. University of the Chinese Academy of Sciences, Beijing 100049, China;
c. The Xi'an Key Laboratory of Pathogenic Microorganism and Tumor Immunity, School of Basic Medicine, Xi'an Medical University, Xi'an 710021, China

Corresponding author: Wei Wang, wangwei@wh.iov.cn
Received Date: 18 January 2024
Accepted Date: 31 May 2024

Abstract

Lassa virus (LASV) is an enveloped, negative-sense RNA virus that causes Lassa hemorrhagic fever. Successful entry of LASV requires the viral glycoprotein 1 (GP1) to undergo a receptor switch from its primary receptor alpha-dystroglycan (α-DG) to its endosomal receptor lysosome-associated membrane protein 1 (LAMP1). A conserved histidine triad in LASV GP1 has been reported to be responsible for receptor switch. To test the hypothesis that other non-conserved residues also contribute to receptor switch, we constructed a series of mutant LASV GP1 proteins and tested them for binding to LAMP1. Four residues, L84, K88, L107, and H170, were identified as critical for receptor switch. Substituting any of the four residues with the corresponding lymphocytic choriomeningitis virus (LCMV) residue (L84 N, K88E, L10F, and H170S) reduced the binding affinity of LASV GP1 for LAMP1. Moreover, all mutations caused decreases in glycoprotein precursor (GPC)-mediated membrane fusion at both pH 4.5 and 5.2. The infectivity of pseudotyped viruses bearing either GPCL84N or GPCK88E decreased sharply in multiple cell types, while L107F and H170S had only mild effects on infectivity. Using biolayer light interferometry assay, we found that all four mutants had decreased binding affinity to LAMP1, in the order of binding affinity being L84 N > L107F > K88E > H170S. The four amino acid loci identified for the first time in this study have important reference significance for the in-depth investigation of the mechanism of receptor switching and immune escape of LASV occurrence and the development of reserve anti-LASV infection drugs.
- Lassa virus (LASV)
- , Lysosome-associated membrane protein 1 (LAMP1)
- , Glycoprotein
- , Receptor switch
- , Membrane fusion

References
1. Acciani M, Alston JT, Zhao G, Reynolds H, Ali AM, Xu B, Brindley MA. 2017. Mutational analysis of Lassa virus glycoprotein highlights regions required for alpha-dystroglycan utilization. J Virol, 91, e00574-17.
2. Cao J, Zhang G, Zhou M, Liu Y, Xiao G, Wang W. 2021. Characterizing the Lassa virus envelope glycoprotein membrane proximal external region for its role in fusogenicity. Virol Sin, 36: 273-280.
3. Carette JE, Raaben M, Wong AC, Herbert AS, Obernosterer G, Mulherkar N, Kuehne AI, Kranzusch PJ, Griffin AM, Ruthel G, Cin PD, Dye JM, Whelan SP, Chandran K, Brummelkamp TR. 2011. Ebola virus entry requires the cholesterol transporter Niemann-Pick C1. Nature, 477: 340-343.
4. Chukwudozie OS. 2020. The function annotations of ST3GAL4 in human lamp1 and lassa virus GP-C interaction from the perspective of systems virology. Access Microbiol, 2: acmi000146.
5. Cohen-Dvashi H, Kilimnik I, Diskin R. 2018. Structural basis for receptor recognition by Lujo virus. Nat Microbiol, 3: 1153-1160.
6. Cohen-Dvashi H, Cohen N, Israeli H, Diskin R. 2015. Molecular mechanism for LAMP1 recognition by Lassa virus. J Virol, 89: 7584-7592.
7. Cohen-Dvashi H, Israeli H, Shani O, Katz A, Diskin R. 2016. Role of LAMP1 binding and ph sensing by the spike complex of Lassa virus. J Virol, 90: 10329-10338.
8. Dong S, Mao W, Liu Y, Jia X, Zhang Y, Zhou M, Hou Y, Xiao G, Wang W. 2023. Deletion of the first glycosylation site promotes Lassa virus glycoprotein-mediated membrane fusion. Virol Sin, 38: 380-386.
9. Enria DA, Briggiler AM, Fernandez NJ, Levis SC, Maiztegui JI. 1984. Importance of dose of neutralising antibodies in treatment of argentine haemorrhagic fever with immune plasma. Lancet, 2: 255-256.
10. Goeijenbier M, Wagenaar J, Goris M, Martina B, Henttonen H, Vaheri A, Reusken C, Hartskeerl R, Osterhaus A, Van Gorp E. 2013. Rodent-borne hemorrhagic fevers: Under-recognized, widely spread and preventable- epidemiology, diagnostics and treatment. Crit Rev Microbiol, 39: 26-42.
11. Hulseberg CE, Feneant L, Szymanska KM, White JM. 2018. LAMP1 increases the efficiency of Lassa virus infection by promoting fusion in less acidic endosomal compartments. mBio, 9, e01818-17.
12. Israeli H, Cohen-Dvashi H, Shulman A, Shimon A, Diskin R. 2017. Mapping of the Lassa virus LAMP1 binding site reveals unique determinants not shared by other old world arenaviruses. PLoS Pathog, 13: e1006337.
13. Jae LT, Brummelkamp TR. 2015. Emerging intracellular receptors for hemorrhagic fever viruses. Trends Microbiol, 23: 392-400.
14. Jae LT, Raaben M, Herbert AS, Kuehne AI, Wirchnianski AS, Soh TK, Stubbs SH, Janssen H, Damme M, Saftig P, Whelan SP, Dye JM, Brummelkamp TR. 2014. Virus entry. Lassa virus entry requires a trigger-induced receptor switch. Science, 344: 1506-1510.
15. Kumar S, Sarkar A. 2019. Capturing the inherent structural dynamics of the HIV-1 envelope glycoprotein fusion peptide. Nat Commun, 10: 763.
16. Li S, Sun Z, Pryce R, Parsy ML, Fehling SK, Schlie K, Siebert CA, Garten W, Bowden TA, Strecker T, Huiskonen JT. 2016. Acidic pH-induced conformations and LAMP1 binding of the Lassa virus glycoprotein spike. PLoS Pathog, 12: e1005418.
17. Liu Y, Guo J, Cao J, Zhang G, Jia X, Wang P, Xiao G, Wang W. 2021. Screening of botanical drugs against Lassa virus entry. J Virol, 95, e02429-20.
18. Moraz ML, Pythoud C, Turk R, Rothenberger S, Pasquato A, Campbell KP, Kunz S. 2013. Cell entry of Lassa virus induces tyrosine phosphorylation of dystroglycan. Cell Microbiol, 15: 689-700.
19. Nunberg JH, York J. 2012. The curious case of arenavirus entry, and its inhibition. Viruses, 4: 83-101.
20. Oppliger J, Torriani G, Herrador A, Kunz S. 2016. Lassa virus cell entry via dystroglycan involves an unusual pathway of macropinocytosis. J Virol, 90: 6412-6429.
21. Raaben M, Jae LT, Herbert AS, Kuehne AI, Stubbs SH, Chou YY, Blomen VA, Kirchhausen T, Dye JM, Brummelkamp TR, Whelan SP. 2017. NRP2 and CD63 are host factors for lujo virus cell entry. Cell Host Microbe, 22: 688-696 e685.
22. Radoshitzky SR, Bao Y, Buchmeier MJ, Charrel RN, Clawson AN, Clegg CS, DeRisi JL, Emonet S, Gonzalez JP, Kuhn JH, Lukashevich IS, Peters CJ, Romanowski V, Salvato MS, Stenglein MD, de la Torre JC. 2015. Past, present, and future of arenavirus taxonomy. Arch Virol, 160: 1851-1874.
23. Sanchez A, Pifat DY, Kenyon RH, Peters CJ, McCormick JB, Kiley MP. 1989. Junin virus monoclonal antibodies: Characterization and cross-reactivity with other arenaviruses. J Gen Virol, 70: 1125-1132.
24. Shimojima M, Stroher U, Ebihara H, Feldmann H, Kawaoka Y. 2012. Identification of cell surface molecules involved in dystroglycan-independent Lassa virus cell entry. J Virol, 86: 2067-2078.
25. Thomson EC, Rosen LE, Shepherd JG, Spreafico R, da Silva Filipe A, Wojcechowskyj JA, Davis C, Piccoli L, Pascall DJ, Dillen J, Lytras S, Czudnochowski N, Shah R, Meury M, Jesudason N, De Marco A, Li K, Bassi J, O'Toole A, Pinto D, Colquhoun RM, Culap K, Jackson B, Zatta F, Rambaut A, Jaconi S, Sreenu VB, Nix J, Zhang I, Jarrett RF, Glass WG, Beltramello M, Nomikou K, Pizzuto M, Tong L, Cameroni E, Croll TI, Johnson N, Di Iulio J, Wickenhagen A, Ceschi A, Harbison AM, Mair D, Ferrari P, Smollett K, Sallusto F, Carmichael S, Garzoni C, Nichols J, Galli M, Hughes J, Riva A, Ho A, Schiuma M, Semple MG, Openshaw PJM, Fadda E, Baillie JK, Chodera JD, Rihn SJ, Lycett SJ, Virgin HW, Telenti A, Corti D, Robertson DL, Snell G. 2021. Circulating SARS-CoV-2 spike N439K variants maintain fitness while evading antibody-mediated immunity. Cell, 184: 1171-1187.e1120.
26. Wang H, Shi Y, Song J, Qi J, Lu G, Yan J, Gao GF. 2016. Ebola viral glycoprotein bound to its endosomal receptor Niemann-Pick C1. Cell, 164: 258-268.
27. Wang P, Liu Y, Zhang G, Wang S, Guo J, Cao J, Jia X, Zhang L, Xiao G, Wang W. 2018. Screening and identification of Lassa virus entry inhibitors from an FDA-approved drug library. J Virol, 92, e00954-18.
28. Wang W, Zhou Z, Zhang L, Wang S, Xiao G. 2016. Structure-function relationship of the mammarenavirus envelope glycoprotein. Virol Sin, 31: 380-394.
29. York J, Nunberg JH. 2006. Role of the stable signal peptide of Junin arenavirus envelope glycoprotein in pH-dependent membrane fusion. J Virol, 80: 7775-7780.
30. Yun NE, Walker DH. 2012. Pathogenesis of Lassa fever. Viruses, 4: 2031-2048.
31. Zhu X, Liu Y, Guo J, Cao J, Wang Z, Xiao G, Wang W. 2021. Effects of N-linked glycan on Lassa virus envelope glycoprotein cleavage, infectivity, and immune response. Virol Sin, 36: 774-783.
Proportional views