A Comparison of the Nucleotide Sequences of Eastern and Western Equine Encephalomyelitis Viruses with Those of Other Alphaviruses and Related RNA Viruses

Weaver, Scott C.; Hagenbaugh, Amy; Bellew, Liz Anne; Нетесов, С. В.; Volchkov, Viktor E.; Chang, Gwong‐Jen J.; Clarke, David K.; Gousset, Laurent; Scott, Thomas W.; Trent, Dennis W.; Holland, John J.

doi:10.1006/viro.1993.1599

Cited by 80 publications

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“…The data in this report indicate that distinctions exist between the electrophoretic profiles and peptide maps of the E 1 and E2 glycoproteins of NA-and SA-type viruses. This information is consistent with other serological, genetic, antigenic and biological evidence that EEE virus strains are distinguishable on the basis of antigenic serotype (Casals, 1964;Walder et al, 1980;Roehrig et al, 1990;Weaver et al, 1991Weaver et al, , 1994. A marked similarity was found to exist among the polypeptides of the NAserotype isolates examined, regardless of host origin, geographic location, year of isolation or passage history.…”

Section: Discussionsupporting

confidence: 89%

“…Supernatant solutions containing the viral envelope proteins E1 and E2 were diluted 1:1 with 0.4 MNaC1, 0.1 M-sodium succinate pH 5"5 and 0.1% Triton X-100. The El/E2 solution was then loaded onto the column and, after washing with 20 ml of equilibration buffer, the void volume containing the (Weaver et al, 1993(Weaver et al, , 1994 Peptide mapping. Purified preparations of radiolabelled El, E2 and C proteins (2 to 7 ~tg) were diluted to 30 lal with V8 digestion buffer and 2-ME (0-9 M final concentration) and heated to 100 °C for 5 min prior to addition of enzyme.…”

Section: Purification Of Individual Virion Structural Proteinsmentioning

confidence: 99%

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Structural Protein Relationships Among Eastern Equine Encephalitis Viruses

Strizki

Repik

1994

Journal of General Virology

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We have re-evaluated the relationships among the polypeptides of eastern equine encephalitis (EEE) viruses using SDS-PAGE and peptide mapping of individual virion proteins. Four to five distinct polypeptide bands were detected upon SDS-PAGE analysis of viruses: the El, E2 and C proteins normally associated with alphavirus virions, as well as an additional more rapidlymigrating E2-associated protein and a high 3//,. (HMW) protein. In contrast with previous findings by others, the electrophoretic profiles of the virion proteins of EEE viruses displayed a marked correlation with serotype. The protein profiles of the 33 North American (NA)-serotype viruses examined were remarkably homogeneous, with variation detected only in the E1 protein of two isolates, In contrast, considerable heterogeneity was observed in the migration profiles of both the E1 and E2 glycoproteins of the 13 South American (SA)-type viruses examined. Peptide mapping of individual virion proteins using limited proteolysis with Staphylococcus aureus V8 protease confirmed that, in addition to the homogeneity evident among NA-type viruses and relative heterogeneity among SA-type viruses, the E1 and E2 proteins of NA-and SA-serotype viruses exhibited serotype-specific structural variation. The C protein was highly conserved among isolates of both virus serotypes. Endoglycosidase analyses of intact virions did not reveal substantial glycosylation differences between the glycoproteins of NA-and SAserotype viruses. Both the HMW protein and the E2 protein (doublet) of EEE virus appeared to contain, at least in part, high-mannose type N-linked oligosaccharides. No evidence of O-linked glycans was found on either the E1 or the E2 glycoprotein. Despite the observed structural differences between proteins of NAand SA-type viruses, Western blot analyses utilizing polyclonal antibodies indicated that immunoreactive epitopes appeared to be conserved.

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Section: Discussionsupporting

confidence: 89%

Section: Purification Of Individual Virion Structural Proteinsmentioning

confidence: 99%

Structural Protein Relationships Among Eastern Equine Encephalitis Viruses

Strizki

Repik

1994

Journal of General Virology

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“…Comparisons using the amino acid sequences of several alphaviruses has confirmed that the envelope glycoproteins were derived from the SINVlike ancestor, whilst the capsid protein was derived from the EEEV-like ancestor (Levinson et al, 1990). Further phylogenetic analyses and nucleotide sequence comparison data demonstrated that WEEV also acquired the genes for non-structural proteins from the EEEV-like ancestor (Weaver et al, 1993). Whilst several pathogenesis studies have been carried out for both SINV and EEEV (Del Piero et al, 2001;Griffin, 1989;Hirsch & Griffin, 1979;Polo et al, 1988;Sherman & Griffin, 1990), the pathogenicity of WEEV remains relatively undescribed (Forrester et al, 2008;Fraizer et al, 1985;Gardner et al, 2008;Hardy et al, 1997;Julander et al, 2007;Nagata et al, 2006;Zlotnik et al, 1972).…”

Section: Introductionmentioning

confidence: 92%

Virulence variation among isolates of western equine encephalitis virus in an outbred mouse model

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Bosio

Welte

et al. 2009

Journal of General Virology

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Little is known about viral determinants of virulence associated with western equine encephalitis virus (WEEV). Here, we have analysed six North American WEEV isolates in an outbred CD1 mouse model. Full genome sequence analyses showed ¡2.7 % divergence among the six WEEV isolates. However, the percentage mortality and mean time to death (MTD) varied significantly when mice received subcutaneous injections of 10 3 p.f.u. of each virus. Two WEEV strains, McMillan (McM) and Imperial 181 (IMP), were the most divergent of the six in genome sequence; McM caused 100 % mortality by 5 days post-infection, whereas IMP caused no mortality. McM had significantly higher titres in the brain than IMP. Similar differences in virulence were observed when McM and IMP were administered by aerosol, intranasal or intravenous routes. McM was 100 % lethal with an MTD of 1.9 days when 10 3 p.f.u. of each virus was administered by intracerebral inoculation; in contrast, IMP caused no mortality. The presence of IMP in the brains after infection by different routes and the lack of observed mortality confirmed that IMP is neuroinvasive but not neurovirulent. Based on morbidity, mortality, MTD, severity of brain lesions, virus distribution patterns, routes of infection and differences in infection of cultured cells, McM and IMP were identified as high-and low-virulence isolates, respectively.

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“…The structural proteins are encoded in the 3′ third of the genome and are translated from a subgenomic RNA, 26S, resulting in 5 protein products: C, 6K, E1, E2, and E3 (2). Previous sequencing studies analyzed genetic relationships of EEEV strains in the Western Hemisphere and compared strains distributed across widespread geographic regions (3)(4)(5)(6). EEEV has 4 distinct genetic lineages; lineage I consists of highly conserved strains from North America, and lineages II-IV encompass strains from Central and South America (3).…”

Section: E Astern Equine Encephalitis Virus (Eeev; Genus Alphavirusmentioning

confidence: 99%