Novel Flavivirus Attenuation Markers Identified in the Envelope Protein of Alfuy Virus
- PMID: 33498300
- PMCID: PMC7909262
- DOI: 10.3390/v13020147
Novel Flavivirus Attenuation Markers Identified in the Envelope Protein of Alfuy Virus
Abstract
Alfuy (ALFV) is an attenuated flavivirus related to the Murray Valley encephalitis virus (MVEV). We previously identified markers of attenuation in the envelope (E) protein of the prototype strain (ALFV3929), including the hinge region (E273-277) and lack of glycosylation at E154-156. To further determine the mechanisms of attenuation we assessed ALFV3929 binding to glycosaminoglycans (GAG), a known mechanism of flaviviruses attenuation. Indeed, ALFV3929 exhibited reduced binding to GAG-rich cells in the presence of heparin; however, low-passage ALFV isolates were relatively unaffected. Sequence comparisons between ALFV strains and structural modelling incriminated a positively-charged residue (K327) in ALFV3929 as a GAG-binding motif. Substitution of this residue to the corresponding uncharged residue in MVEV (L), using a previously described chimeric virus containing the prM & E genes of ALFV3929 in the backbone of MVEV (MVEV/ALFV-prME), confirmed a role for K327 in enhanced GAG binding. When the wild type residues at E327, E273-277 and E154-156 of ALFV3929 were replaced with the corresponding residues from virulent MVEV, it revealed each motif contributed to attenuation of ALFV3929, with the E327/E273-277 combination most dominant. These data demonstrate that attenuation of ALFV3929 is multifactorial and provide new insights for the rational design of attenuated flavivirus vaccines.
Keywords: Alfuy virus (ALFV); Murray Valley encephali-tis virus (MVEV); attenuation; flavivirus; glycosaminoglycan (GAG).
Conflict of interest statement
The authors declare no conflict of interest.
Figures




Similar articles
-
Determinants of attenuation in the envelope protein of the flavivirus Alfuy.J Gen Virol. 2011 Oct;92(Pt 10):2286-2296. doi: 10.1099/vir.0.034793-0. Epub 2011 Jul 6. J Gen Virol. 2011. PMID: 21733886
-
Mechanism of virulence attenuation of glycosaminoglycan-binding variants of Japanese encephalitis virus and Murray Valley encephalitis virus.J Virol. 2002 May;76(10):4901-11. doi: 10.1128/jvi.76.10.4901-4911.2002. J Virol. 2002. PMID: 11967307 Free PMC article.
-
Biological, antigenic and phylogenetic characterization of the flavivirus Alfuy.J Gen Virol. 2006 Feb;87(Pt 2):329-337. doi: 10.1099/vir.0.81252-0. J Gen Virol. 2006. PMID: 16432019
-
The Japanese encephalitis serological group of flaviviruses: a brief introduction to the group.Curr Top Microbiol Immunol. 2002;267:1-10. doi: 10.1007/978-3-642-59403-8_1. Curr Top Microbiol Immunol. 2002. PMID: 12082984 Review. No abstract available.
-
The emergence of West Nile virus: the Australian connection.Viral Immunol. 2000;13(4):447-61. doi: 10.1089/vim.2000.13.447. Viral Immunol. 2000. PMID: 11192291 Review. No abstract available.
Cited by
-
Glycosaminoglycan binding by arboviruses: a cautionary tale.J Gen Virol. 2022 Feb;103(2):001726. doi: 10.1099/jgv.0.001726. J Gen Virol. 2022. PMID: 35191823 Free PMC article. Review.
-
Attenuating Mutations in Usutu Virus: Towards Understanding Orthoflavivirus Virulence Determinants and Live Attenuated Vaccine Design.Vaccines (Basel). 2025 May 3;13(5):495. doi: 10.3390/vaccines13050495. Vaccines (Basel). 2025. PMID: 40432107 Free PMC article.
-
Evidence of Infection with Zoonotic Mosquito-Borne Flaviviruses in Saltwater Crocodiles (Crocodylus porosus) in Northern Australia.Viruses. 2022 May 21;14(5):1106. doi: 10.3390/v14051106. Viruses. 2022. PMID: 35632847 Free PMC article.
-
Genomic Determinants Potentially Associated with Clinical Manifestations of Human-Pathogenic Tick-Borne Flaviviruses.Int J Mol Sci. 2022 Nov 2;23(21):13404. doi: 10.3390/ijms232113404. Int J Mol Sci. 2022. PMID: 36362200 Free PMC article.
-
Emerging mosquito-borne flaviviruses.mBio. 2024 Dec 11;15(12):e0294624. doi: 10.1128/mbio.02946-24. Epub 2024 Oct 31. mBio. 2024. PMID: 39480108 Free PMC article. Review.
References
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources