Sequence Determination of the Sendai Virus Fusion Protein Gene

SUMMARYAmino acid sequences of fusion (F) proteins of two trypsin-resistant mutants of Sendai virus, TR-2 and TR-5, were deduced from nucleotide analysis of cDNA encoding the F gene and were compared with that of the trypsin-sensitive wild-type Sendai virus. In both mutants, amino acid substitutions were found at residues 116 (Arg ~ Ile), the cleavage site of the F protein, and 109 (Asn --, Asp). Two trypsinsensitive revertants, TSrev-52 and TSrev-58, derived from TR-5 were both activated by trypsin similarly to the wild-type virus and had a single amino acid reversion from Ile to Arg at residue 116, leaving Asp as before at residue 109. These results indicate that the trypsin sensitivity of Sendal virus can be changed by a single amino acid substitution at the cleavage site of the F protein and a mutation from Arg to Ile is responsible for the acquisition of resistance to trypsin.

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confidence: 56%

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confidence: 99%

Single Amino Acid Substitution of Sendai Virus at the Cleavage Site of the Fusion Protein Confers Trypsin Resistance

Itoh

Shibuta

Homma

1987

“…2 shows a hydropathy profile of the NDV F amino acid sequence, in which the corresponding three regions of high hydrophobicity are indicated. In contrast to the NDV HN amino acid sequence , the F sequence is markedly more hydrophobic than the average, On the basis of the similar high hydrophobicity of the F polypeptide of Sendal virus, it has been suggested that regions of the F glycoprotein in addition to the N terminus of F1 may be capable of hydrophobic interactions with membranes during the fusion process (Blumberg et al, 1985).…”

Section: Resultsmentioning

confidence: 97%

“…Modifications to F0 include cleavage of the signal sequence (Blumberg et al, 1985), glycosylation (Mountcastle et al, 1971), fatty acid acylation (Schmidt, 1982;Chatis & Morrison, 1982) and possibly rearrangements of intramolecular disulphide bonds (McGinnes et al, 1985). Modifications to F 2 include blockage of the N terminus (Scheid et al, 1978) and trimming of the C terminus by carboxypeptidase (Kohama et al, 1981), Determination of the amino acid sequence of F of NDV may suggest the locations of some of these processing events in the F polypeptide chain.…”

Section: Introductionmentioning

confidence: 99%

Nucleotide Sequence of the Gene Encoding the Fusion Glycoprotein of Newcastle Disease Virus

Chambers

Millar

Emmerson

1986

SUMMARYThe nucleotide sequence of the gene encoding the fusion (F) glycoprotein of the Beaudette C strain of Newcastle disease virus (NDV) has been determined from cDNA clones obtained from virion RNA. The gene is 1792 nucleotides long, including mRNA start and polyadenylation signals typical of paramyxoviruses. The single open reading frame encodes a polypeptide of 553 amino acids, with a predicted molecular weight of 59042. The F polypeptide has three regions of high hydrophobicity: an Nterminal signal peptide, the N terminus of F~ (known from protein sequencing) and a C-terminal membrane-spanning region by which the F glycoprotein is anchored to the membrane. The cleavage site of F0 is located in a highly basic region of the F polypeptide. Five potential asparagine-linked glycosylation sites are present in the amino acid sequence, of which one is in Fz and the others in F~. Comparison of the NDV F amino acid sequence to those from other paramyxoviruses reveals homology to Sendai virus, simian virus 5 and human respiratory syncytial virus. There is also limited homology between the N terminus ofF1 of NDV and the N termini of HA2 of influenza viruses. Post-translational modifications of the NDV F polypeptide are discussed in the light of information provided by the amino acid sequence.

“…The [35S]methionine-labelled X protein was found to comigrate with the [3H]glucosaminelabelled F2 protein of Sendai virus during SDS-PAGE [the F2 protein does not contain any methionine residues (Blumberg et al, 1985)]. However, unlike the F2 protein, the X protein could not be labelled with glucosamine (not shown).…”

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confidence: 98%

Identification of an Additional Sendai Virus Non-structural Protein Encoded by the P/C mRNA

Curran¹,

Kolakofsky

1987

Self Cite

SUMMARYPeptide antiserum and monoclonal antibodies have detected a previously unrecognized small non-structural protein in Sendai virus-infected cells. This protein, designated X, appears to represent the C-terminal 95 amino acids of the P protein reading frame. The X protein appears to be almost as abundant as the P protein on a molar basis in vivo. No evidence of a precursor-product relationship between the X and P proteins could be found.Sendai virus, a member of the parainfluenza virus genus of the paramyxovirus family, encodes its entire genetic information within a non-segmented 15 kb negative strand (-) RNA genome (Kingsbury, 1974;Kolakofsky et al., 1974). Upon infection, this genetic information is expressed via positive-sense mRNAs transcribed from the-genome by the virion-associated polymerase. These transcripts are largely monocistronic, encoding the NP, M, F, HN and L proteins. The P mRNA, however, has been demonstrated to be polycistronic. In addition to encoding the 568 amino acid (aa) P protein, it also codes for a pair of non-structural proteins, C (204 aa) and C' (181 aa) (Curran et al., 1986;Dethlefsen & Kolakofsky, 1983;Giorgi et al., 1983) which represent a C-coterminal nested set and are derived from an overlapping open reading frame (ORF) (see Fig. 1). The C overlapping ORF has also been conserved in the closely related human parainfluenza virus 3 (Spriggs & Collins, 1986a) as well as the two morbilliviruses examined to date, measles virus and canine distemper virus (Barrett et al., 1985;Bellini et al., 1985). Curiously, in the case of the parainfluenza viruses simian virus 5, Newcastle disease virus and mumps virus, non-structural proteins are again encoded within the P mRNA, but here these non-structural proteins appear to share tryptic peptides with the P protein and are therefore thought to be encoded by the same ORF (Collins et al., 1982;Herrler & Compans, 1982;Paterson et al., 1984). Only in the case of respiratory syncytial virus, a member of the more distinctly related pneumovirus genus, is the P mRNA thought to be monocistronic (Collins et al., 1984).Recently, Herman (1986) has found that the NS mRNA of the rhabdovirus vesicular stomatitis virus (VSV), which is thought to be the VSV equivalent of the parainfluenza virus P mRNA, also codes for a second small protein, representing the C-terminal 62 aa of the 265 aa long NS protein. Since the Sendal virus P protein and the VSV NS protein have several characteristics in common, and antiserum specific for the C-terminal t6 aa of the Sendai virus P protein is available, we examined whether the same also applies to Sendai virus.Sendai virus (Harris strain)-infected BHK cells were labelled for 2 h at 18 h post-infection, when marked cytopathic effects are apparent, as well as a parallel uninfected culture. Under these conditions, host protein synthesis has largely been displaced by the viral protein synthesis and the virus structural proteins and the non-structural C protein can clearly be detected above the remaining cellular background ...