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Volume 104, Issue 12

Internalization of rabies virus glycoprotein differs between pathogenic and attenuated virus strains Open Access

Abstract

The zoonotic rabies virus (RABV) is a non-segmented negative-sense RNA virus classified within the family , and is the most common aetiological agent responsible for fatal rabies disease. The RABV glycoprotein (G) forms trimeric spikes that protrude from RABV virions and mediate virus attachment, entry and spread, and is a major determinant of RABV pathogenesis. A range of RABV strains exist that are highly pathogenic in part due to their ability to evade host immune detection. However, some strains are disease-attenuated and can be cleared by host defences. A detailed molecular understanding of how strain variation relates to pathogenesis is currently lacking. Here, we reveal key differences in the trafficking profiles of RABV-G proteins from the challenge virus standard strain (CVS-11) and a highly attenuated vaccine strain SAD-B19 (SAD). We show that CVS-G traffics to the cell surface and undergoes rapid internalization through both clathrin- and cholesterol-dependent endocytic pathways. In contrast, SAD-G remains resident at the plasma membrane and internalizes at a significantly slower rate. Through engineering hybrids of CVS-G and SAD-G, we show that the cytoplasmic tail of CVS-G is the key determinant of these different internalization profiles. Alanine scanning further revealed that mutation of Y497 in CVS-G (H497 in SAD-G) could reduce the rate of internalization to SAD-G levels. Together, these data reveal new phenotypic differences between CVS-G and SAD-G proteins that may contribute to altered pathogenicity.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): attenuated vaccine strain SAD-B19 , challenge virus standard (CVS) strain , internalization, trafficking , rabies virus and rabies virus glycoprotein
© 2023 The Authors
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License. This article was made open access via a Publish and Read agreement between the Microbiology Society and the corresponding author’s institution.
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2023-12-08
2024-05-08
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References

  1. World Health Organization WHO Expert Consultation on Rabies: Third Report World Health Organization; 2018
     [Google Scholar]
  2. Meslin F-X. Rabies as a traveler-s risk, especially in high-endemicity areas. J Travel Med 2008; 12:S30–S40 [View Article]
     [Google Scholar]
  3. Hampson K, Dobson A, Kaare M, Dushoff J, Magoto M et al. Rabies exposures, post-exposure prophylaxis and deaths in a region of endemic canine rabies. PLoS Negl Trop Dis 2008; 2:e339 [View Article] [PubMed]
     [Google Scholar]
  4. Knobel DL, Cleaveland S, Coleman PG, Fèvre EM, Meltzer MI et al. Re-evaluating the burden of rabies in Africa and Asia. Bull World Health Organ 2005; 83:360–368 [PubMed]
     [Google Scholar]
  5. Fisher CR, Streicker DG, Schnell MJ. The spread and evolution of rabies virus: conquering new frontiers. Nat Rev Microbiol 2018; 16:241–255 [View Article] [PubMed]
     [Google Scholar]
  6. Davis BM, Rall GF, Schnell MJ. Everything you always wanted to know about rabies virus (but were afraid to ask). Annu Rev Virol 2015; 2:451–471 [View Article] [PubMed]
     [Google Scholar]
  7. Guo Y, Duan M, Wang X, Gao J, Guan Z et al. Early events in rabies virus infection-attachment, entry, and intracellular trafficking. Virus Res 2019; 263:217–225 [View Article] [PubMed]
     [Google Scholar]
  8. Miyamoto K, Matsumoto S. Comparative studies between pathogenesis of street and fixed rabies infection. J Exp Med 1967; 125:447–456 [View Article] [PubMed]
     [Google Scholar]
  9. Murphy FA. Rabies pathogenesis. Arch Virol 1977; 54:279–297 [View Article] [PubMed]
     [Google Scholar]
  10. Sarmento L, Li X, Howerth E, Jackson AC, Fu ZF. Glycoprotein-mediated induction of apoptosis limits the spread of attenuated rabies viruses in the central nervous system of mice. J Neurovirol 2005; 11:571–581 [View Article] [PubMed]
     [Google Scholar]
  11. Potratz M, Zaeck L, Christen M, Te Kamp V, Klein A et al. Astrocyte infection during rabies encephalitis depends on the virus strain and infection route as demonstrated by novel quantitative 3D analysis of cell tropism. Cells 2020; 9:412 [View Article] [PubMed]
     [Google Scholar]
  12. Morimoto K, Foley HD, McGettigan JP, Schnell MJ, Dietzschold B. Reinvestigation of the role of the rabies virus glycoprotein in viral pathogenesis using a reverse genetics approach. J Neurovirol 2000; 6:373–381 [View Article] [PubMed]
     [Google Scholar]
  13. Faber M, Pulmanausahakul R, Nagao K, Prosniak M, Rice AB et al. Identification of viral genomic elements responsible for rabies virus neuroinvasiveness. Proc Natl Acad Sci U S A 2004; 101:16328–16332 [View Article] [PubMed]
     [Google Scholar]
  14. Li C, Zhang H, Ji L, Wang X, Wen Y et al. Deficient incorporation of rabies virus glycoprotein into virions enhances virus-Iinduced immune evasion and viral pathogenicity. Viruses 2019; 11:218 [View Article] [PubMed]
     [Google Scholar]
  15. Yang Y, Huang Y, Gnanadurai CW, Cao S, Liu X et al. The inability of wild-type rabies virus to activate dendritic cells is dependent on the glycoprotein and correlates with its low level of the de novo-synthesized leader RNA. J Virol 2015; 89:2157–2169 [View Article] [PubMed]
     [Google Scholar]
  16. Nitschel S, Zaeck LM, Potratz M, Nolden T, Te Kamp V et al. Point mutations in the glycoprotein ectodomain of field rabies viruses mediate cell culture adaptation through improved virus release in a host cell dependent and independent manner. Viruses 2021; 13:1989 [View Article] [PubMed]
     [Google Scholar]
  17. Finke S, Conzelmann K-K. Replication strategies of rabies virus. Virus Res 2005; 111:120–131 [View Article] [PubMed]
     [Google Scholar]
  18. Scrima N, Le Bars R, Nevers Q, Glon D, Chevreux G et al. Rabies virus P protein binds to TBK1 and interferes with the formation of innate immunity-related liquid condensates. Cell Rep 2023; 42:111949 [View Article] [PubMed]
     [Google Scholar]
  19. Yin K, Li Y, Ma Z, Yang Y, Zhao H et al. SNAP25 regulates the release of the rabies virus in nerve cells via SNARE complex-mediated membrane fusion. Vet Microbiol 2020; 245:108699 [View Article] [PubMed]
     [Google Scholar]
  20. Leonard D, Hayakawa A, Lawe D, Lambright D, Bellve KD et al. Sorting of EGF and transferrin at the plasma membrane and by cargo-specific signaling to EEA1-enriched endosomes. J Cell Sci 2008; 121:3445–3458 [View Article] [PubMed]
     [Google Scholar]
  21. Shakin-Eshleman SH, Remaley AT, Eshleman JR, Wunner WH, Spitalnik SL. N-linked glycosylation of rabies virus glycoprotein. Individual sequons differ in their glycosylation efficiencies and influence on cell surface expression. J Biol Chem 1992; 267:10690–10698 [PubMed]
     [Google Scholar]
  22. Yamada K, Noguchi K, Nonaka D, Morita M, Yasuda A et al. Addition of a single N-glycan to street rabies virus glycoprotein enhances virus production. J Gen Virol 2013; 94:270–275 [View Article] [PubMed]
     [Google Scholar]
  23. Seif I, Coulon P, Rollin PE, Flamand A. Rabies virulence: effect on pathogenicity and sequence characterization of rabies virus mutations affecting antigenic site III of the glycoprotein. J Virol 1985; 53:926–934 [View Article] [PubMed]
     [Google Scholar]
  24. Vigerust DJ, Shepherd VL. Virus glycosylation: role in virulence and immune interactions. Trends Microbiol 2007; 15:211–218 [View Article] [PubMed]
     [Google Scholar]
  25. Yamada K, Park C-H, Noguchi K, Kojima D, Kubo T et al. Serial passage of a street rabies virus in mouse neuroblastoma cells resulted in attenuation: potential role of the additional N-glycosylation of a viral glycoprotein in the reduced pathogenicity of street rabies virus. Virus Res 2012; 165:34–45 [View Article] [PubMed]
     [Google Scholar]
  26. Yamada K, Noguchi K, Nishizono A. Efficient N-glycosylation at position 37, but not at position 146, in the street rabies virus glycoprotein reduces pathogenicity. Virus Res 2014; 179:169–176 [View Article] [PubMed]
     [Google Scholar]
  27. Ochsenbauer C, Dubay SR, Hunter E. The Rous sarcoma virus Env glycoprotein contains a highly conserved motif homologous to tyrosine-based endocytosis signals and displays an unusual internalization phenotype. Mol Cell Biol 2000; 20:249–260 [View Article] [PubMed]
     [Google Scholar]
  28. Favoreel HW, Van Minnebruggen G, Nauwynck HJ, Enquist LW, Pensaert MB. A tyrosine-based motif in the cytoplasmic tail of pseudorabies virus glycoprotein B is important for both antibody-induced internalization of viral glycoproteins and efficient cell-to-cell spread. J Virol 2002; 76:6845–6851 [View Article] [PubMed]
     [Google Scholar]
  29. Ilinskaya A, Heidecker G, Derse D. Opposing effects of a tyrosine-based sorting motif and a PDZ-binding motif regulate human T-lymphotropic virus type 1 envelope trafficking. J Virol 2010; 84:6995–7004 [View Article] [PubMed]
     [Google Scholar]
  30. Royle SJ, Bobanović LK, Murrell-Lagnado RD. Identification of a non-canonical tyrosine-based endocytic motif in an ionotropic receptor. J Biol Chem 2002; 277:35378–35385 [View Article] [PubMed]
     [Google Scholar]
  31. Conzelmann KK, Cox JH, Schneider LG, Thiel HJ. Molecular cloning and complete nucleotide sequence of the attenuated rabies virus SAD B19. Virology 1990; 175:485–499 [View Article] [PubMed]
     [Google Scholar]
  32. Reardon TR, Murray AJ, Turi GF, Wirblich C, Croce KR et al. Rabies virus CVS-N2c(ΔG) strain enhances retrograde synaptic transfer and neuronal viability. Neuron 2016; 89:711–724 [View Article] [PubMed]
     [Google Scholar]
  33. Wickersham IR, Finke S, Conzelmann K-K, Callaway EM. Retrograde neuronal tracing with a deletion-mutant rabies virus. Nat Methods 2007; 4:47–49 [View Article] [PubMed]
     [Google Scholar]
  34. Orbanz J, Finke S. Generation of recombinant European bat lyssavirus type 1 and inter-genotypic compatibility of lyssavirus genotype 1 and 5 antigenome promoters. Arch Virol 2010; 155:1631–1641 [View Article] [PubMed]
     [Google Scholar]
  35. Dwane S, Durack E, Kiely PA. Optimising parameters for the differentiation of SH-SY5Y cells to study cell adhesion and cell migration. BMC Res Notes 2013; 6:366 [View Article] [PubMed]
     [Google Scholar]
  36. Müller T, Dietzschold B, Ertl H, Fooks AR, Freuling C et al. Development of a mouse monoclonal antibody cocktail for post-exposure rabies prophylaxis in humans. PLoS Negl Trop Dis 2009; 3:e542 [View Article] [PubMed]
     [Google Scholar]
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Internalization of rabies virus glycoprotein differs between pathogenic and attenuated virus strains
J. Gen. Virol. 104, 001935 (2023); https://doi.org/10.1099/jgv.0.001935
/content/journal/jgv/10.1099/jgv.0.001935
/content/journal/jgv/10.1099/jgv.0.001935
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