Characterization of the 10 proteins of human respiratory syncytial virus: Identification of a fourth envelope-associated protein

Huang, Yung T.; Collins, Peter L.; Wertz, Gail W.

doi:10.1016/0168-1702(85)90246-1

Cited by 126 publications

(98 citation statements)

References 28 publications

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“…Previously, RSV strain A2 virions that had been purified on sucrose gradients were shown by SDS-PAGE to contain SHo protein, albeit in amounts that were low relative to the other RSV structural proteins and could be detected only with long film exposures (Huang et al, 1985). Factors that complicate the identification of RSV structural proteins include the low yield of released virus in tissue culture, the lability of the virion and the tendency of released virions to be contaminated with intracellular proteins and cellular membranes.…”

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confidence: 99%

“…One of the 10 RSV proteins is a small (64 amino acids for strain A2) hydrophobic species that is called 1A or SH (Collins et al, 1984;Huang et al, 1985;Olmsted & Collins, 1989). The nucleotide and amino acid sequence of the SH mRNA and protein of RSV strain A2 of RSV antigenic subgroup A were determined by cDNA sequencing .…”

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confidence: 99%

“…At 12 h post-infection actinomycin D was added to 2 ~tg/ml, at 17 h post-infection the cells were labelled by incubation with 50pCi/ml [asS]methionine for 1 h with 2~tg/ml actinomycin D and immediately following the labelling period lysates were prepared for SDS-PAGE. Additional cultures were infected in parallel, were incubated without actinomycin D, and were labelled with 20 pCi/ml [35S]methionine from 8 to 26 h post-infection; The supernatant fluids were collected, adjusted to contain 0-1 M-magnesium sulphate and 50 mM-HEPES pH 7.5, clarified, and subjected to two cycles of centrifugation on discontinuous sucrose gradients (Huang et al, 1985) in order to band and partially purify the supernatant RSV. The virus was solubilized in immunoprecipitation buffer (Olmsted & Collins, 1989), clarified and either (lane V) adjusted to the composition of SDS-PAGE sample buffer and analysed directly or (lane SH) following immunoprecipitation with antiserum specific to the C terminus of the SH protein.…”

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confidence: 99%

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The small hydrophobic protein of human respiratory syncytial virus: comparison between antigenic subgroups A and B

Collins

Olmsted

Johnson

1990

Journal of General Virology

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The small hydrophobic protein of human respiratory syncytial virus: comparison between antigenic subgroups A and B

Collins

Olmsted

Johnson

1990

Journal of General Virology

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“…Unlike other paramyxoviruses, RS virus possesses two nonglycosylated proteins associated with the virion membrane : the matrix (M) protein and the 22K protein (Wunner & Pringle, 1976;Peeples & Levine, 1979;Huang et al, 1984Huang et al, , 1985. The 22K protein (which migrates with a relative molecular weight of 23 000 in gels run in our laboratory; Routledge et al, 1985) is relatively hydrophilic and is the most basic of the proteins produced in cells as a result of RS virus infection (Dubovi, 1982), but its function is unknown.…”

Section: Discussionmentioning

confidence: 99%

Expression of the Respiratory Syncytial Virus 22K Protein on the Surface of Infected HeLa Cells

Routledge¹,

Willcocks

Morgan³

et al. 1987

Journal of General Virology

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“…They are enveloped, single-stranded, nonsegmented, negativesense RNA viruses. 11 The genome encodes 10 viral proteins: 1B, 1C, N, P, M, SH, M2, L, the major glycoprotein (G), and the fusion (F) protein. 3,8 The G and F proteins are present in the membrane of infected cells and in the virus envelope.…”

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Validation of Synthetic Peptide Enzyme Immunoassays in Differentiating Two Subgroups of Ruminant Respiratory Syncytial Virus

2001

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Abstract. Subgroup-specific peptide-based enzyme immunoassays from each respective G-glycoprotein of the ovine and the bovine respiratory syncytial virus (RSV) were developed to detect RSV-specific IgG responses in cattle. Antigenic peptides from the respective G-glycoprotein were identified from the extracellular central hydrophobic region (amino acids 158-189) located between 2 mucin-rich regions. These antigenic peptides identified by epitope mapping from each G-glycoprotein were synthesized and used to develop the subgroupspecific enzyme immunoassays. The negative cutoff for each enzyme immunoassay was established as the mean optical density of indirect immunofluorescent antibody-negative bovine sera plus 3 SDs. The sensitivity (82.9%) and specificity (100%) of the bovine enzyme immunoassay and the specificity (95.8%) of the ovine enzyme immunoassay were determined by comparison with indirect immunofluorescence (used as the ''gold standard''). The negative and positive predictive values were calculated for each assay. The presence of serum antibody to ovine RSV in cattle implies that this virus infects cattle and may contribute to the pathogenesis of bovine respiratory disease.Economic losses due to respiratory disease in the US cattle industry approximate $1 billion annually. 10 Respiratory syncytial virus (RSV) has been isolated from people (human RSV [HRSV]) and from several animal species, including cattle (bovine RSV [BRSV]), sheep (ovine RSV [ORSV]), and goats (caprine RSV). These viruses are members of the genus Pneumovirus in the family Paramyxoviridae. They are enveloped, single-stranded, nonsegmented, negativesense RNA viruses. 11 The genome encodes 10 viral proteins: 1B, 1C, N, P, M, SH, M2, L, the major glycoprotein (G), and the fusion (F) protein. 3,8 The G and F proteins are present in the membrane of infected cells and in the virus envelope. 16 The attachment protein G has been the most variable antigen among viral strains. 2 The proposed 3-dimensional model for the Gglycoprotein of BRSV contains a hydrophobic globular region (residues 158-189) bordered by 2 hydrophilic densely carbohydrate-coated mucin-like regions. 15 The G-glycoprotein gene from an ORSV strain has been cloned and sequenced and had greater homology to the BRSV gene (and its encoded protein) than to the HRSV gene (and its encoded protein). 2,4 However, the degree of base sequence differences between BRSV and ORSV suggests that 2 subgroups of an ungulate RSV exist. 2,15 Recent RNase protection assays, Received for publication October 26, 1999. reverse transcription polymerase chain reaction amplification studies, and a phosphoprotein profile analysis support the existence of 2 ruminant RSV subgroups. 1,9,17,18 Current knowledge concerning interspecies transmission of RSV strains is limited. Antibodies to BRSV were found in cattle, goats, and horses but were not detected in sheep. 14,20 Recently, serologic evidence of RSV infection in swine has been reported. 5 Respiratory syncytial virus is recognized as an important pathog...

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Characterization of the 10 proteins of human respiratory syncytial virus: Identification of a fourth envelope-associated protein

Cited by 126 publications

References 28 publications

The small hydrophobic protein of human respiratory syncytial virus: comparison between antigenic subgroups A and B

The small hydrophobic protein of human respiratory syncytial virus: comparison between antigenic subgroups A and B

Expression of the Respiratory Syncytial Virus 22K Protein on the Surface of Infected HeLa Cells

Validation of Synthetic Peptide Enzyme Immunoassays in Differentiating Two Subgroups of Ruminant Respiratory Syncytial Virus

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