Structural Polypeptides of Coronavirus IBV

Cavanagh, David

doi:10.1099/0022-1317-53-1-93

Cited by 93 publications

(87 citation statements)

References 28 publications

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“…The radiolabelling of virus with 35 S-methionine in de-embryonated eggs, the purification of the virus and its analysis by SDS-polyacrylamide gel electrophoresis have been described (Cavanagh, 1981(Cavanagh, ,1983.…”

Section: Radiolabelled Virus Proteinsmentioning

confidence: 99%

Confirmation of the presence of avian infectious bronchitis virus (IBV) using cloned DNA complementary to the 3’ terminus of the IBV genome

1984

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SUMMARYA hybridisation test has been developed to aid the identification of virus isolates as avian infectious bronchitis virus (IBV) when serum neutralisation tests have proved inadequate. In this test nucleic acid prepared from four recent virus isolates suspected of being IBV hybridised with a 32 P-radiolabelled cloned cDNA (complementary DNA) probe complementary to RNA from IBV strain Beaudette. Since these strains represent one or more new serotypes they had not been identified by neutralisation tests using antisera to known IBV serotypes. Polyacrylamide gel electrophoresis of the isolates radiolabelled in de-embryonated eggs showed that the viruses had proteins characteristic of IBV. Nine other IBV strains, representing five serotypes, also hybridised with the probe. (Hopkins, 1974) infectious bronchitis in the United Kingdom is largely controlled by live vaccines of the Massachusetts serotype. However, Cook (1983) has isolated five strains of virus from chickens of five flocks which had been vaccinated against IBV. Although coronavirus-like particles were observed by electron microscopy four strains appeared to be serologically distinct from existing IBV serotypes since they were not neutralised by antisera prepared against strains of five known serotypes (Cook, 1983). It was clearly desirable to confirm that the recent isolates were IBV. To this end a test has been developed for IBV which employs a 32 P-labelled probe prepared from a plasmid which contains cloned DNA complementary (cDNA) to a nucelotide sequence near the 3' end of the genomic RNA of IBV strain Beaudette. Tests of this type have been shown to be highly specific under appropriate conditions (Howley et al., 1979). RNA from crude preparations of putative and known IBV isolates is bound to nitrocellulose filters and then exposed to the probe. Binding of the probe to the filters is readily detected by auto radiography. The test can be used to help identify an isolate as IBV. INTRODUCTION Although infectious bronchitis virus (IBV) exhibits extensive antigenic variation

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Section: Radiolabelled Virus Proteinsmentioning

confidence: 99%

Confirmation of the presence of avian infectious bronchitis virus (IBV) using cloned DNA complementary to the 3’ terminus of the IBV genome

1984

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“…Radiolabelled preparations of each strain were rnade in pairs of de-embryonated eggs using 250 juCi of 35S-methionine (Amersham International, UK) per egg and purified by ammonium sulphate precipitation followed by sucrose gradient sedimentation (Cavanagh, 1981). Virus in the density range 1.15 to 1.21 g/ml was pelleted by centrifugation for 3 hours at 90Kg max at 4°C in a 6 x 14 ml swing-out rotor (MSE).…”

Section: Preparation Of Radiolabelled Virusmentioning

confidence: 99%

“…IBV has two glycoproteins (Cavanagh, 1981). The surface projection or spike (S) glycoprotein comprises two glycopolypeptides S1 and S2 derived by cleavage of a precursor glycopolypeptide (Stern and Sefton, 1982a;Cavanagh, 1983b, c;Cavanagh et al, 1986a).…”

Section: Introductionmentioning

confidence: 99%

Coronavirus IBV: Relationships among recent European isolates studied by limited proteolysis of the Virion Glycopolypeptides

Cavanagh

Davis

1987

Avian Pathology

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SUMMARYAfter the year 1978, strains of avian infectious bronchitis virus (IBV) were isolated in the Netherlands and the UK which were assigned by neutralisation tests to four new serotypes (arbitrarily designated A, B, C and D in this communication) distinct from the long recognised US serotypes. We have labelled, with 35S-methionine, the structural polypeptides of 12 of the European isolates during growth in de-embryonated eggs. The S2, spike-anchoring, polypeptide of isolates of all four serotypes had a molecular weight of 87 000 (87K) whereas the US IBV-M41 strain had S2 of 84K. When the isolates were grouped according to the molecular weight of the other spike glycopolypeptide, S1, and the membrane (M) glycopolypeptide four groups emerged which corresponded to the serotypes based on neutralisation tests. Serotypes A (isolates D207, 1, 5 and 6) and B (isolates 7, 8 and D3896) had S1 of 91.5K while serotypes C (isolates 9 and D3128) and D (isolates D212 and D1466) had S1 of 90K. The M of serotypes A and D had a molecular weight of 30K while that of serotypes B and C was 27K. Chymotrypsin and protease V8 were used for limited proteolysis, the hydrolysis of S1 giving the greatest discrimination between serotypes. After proteolysis of S1 the UK isolates of serotype A gave identical profiles which were very similar to the Dutch serotype A isolate D207. Isolates of serotype B gave S1 profiles very similar to those of serotype A, as did D274 which is serologically related to both groups A and B. Serotypes C and D were distinguishable from each other and from serotypes A and B on the basis of the S1 peptide profiles. These results strengthen the view that the UK and Dutch isolates are closely related and that serotypes A and B are more closely related to each other than to either group C or D serotypes.

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“…The E2 glycoprotein (also designated as the spike or S protein) is observed as an uncleaved form of about 170K to 200K and/or cleaved forms of approximately 85K to 100K; the latter forms may either comigrate or migrate as separate entities on polyacrylamide gels (Sturman et al, 1985 ;Cavanagh, 1981 ;Stern & Sefton, 1982). Thus the 5' half of the E2 gene is known as the S1 region and the 3' half is known as the $2 region, yielding the S1 and $2 polypeptides, respectively, after proteolytic cleavage (Binns et aL, 1985;Cavanagh et al, 1986).…”

Section: Introductionmentioning

confidence: 99%

Mapping of Neutralizing Epitopes to Fragments of the Bovine Coronavirus E2 Protein by Proteolysis of Antigen-Antibody Complexes

Deregt

Parker²,

Cox

et al. 1989

Journal of General Virology

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SUMMARYNeutralizing antigenic domains on bovine coronavirus gpl00/E2 were mapped to fragments of this protein by proteolytic cleavage and fragment analysis. The procedure involved analysis of fragments generated after incubation of E2-monoclonal antibody complexes with various proteases. The smallest antibody-bound fragments obtained were a 50K fragment following Staphylococcus aureus V8 protease and submaxillary protease digestion, and a 37K fragment following trypsin digestion. Trypsin also produced a transient antibody-bound 50K fragment. A 40K fragment which was not bound by antibody was observed following digestions with all three proteases. The 50K fragments generated by V8, submaxillary protease and trypsin comigrated on gels and displayed the same altered mobility under non-reducing conditions, suggesting identity of these fragments and indicating the presence of disulphide linkages in these fragments. The 40K fragments generated by these three enzymes also comigrated and displayed the same altered mobility under non-reducing conditions. The 37K trypsin fragment contained both neutralizing domains, A and B.

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Structural Polypeptides of Coronavirus IBV

Cited by 93 publications

References 28 publications

Confirmation of the presence of avian infectious bronchitis virus (IBV) using cloned DNA complementary to the 3’ terminus of the IBV genome

Confirmation of the presence of avian infectious bronchitis virus (IBV) using cloned DNA complementary to the 3’ terminus of the IBV genome

Coronavirus IBV: Relationships among recent European isolates studied by limited proteolysis of the Virion Glycopolypeptides

Mapping of Neutralizing Epitopes to Fragments of the Bovine Coronavirus E2 Protein by Proteolysis of Antigen-Antibody Complexes

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