Cloning and sequencing of the matrix protein (M) gene of turkey rhinotracheitis virus reveal a gene order different from that of respiratory syncytial virus.
Several biochemical properties and the sequence of the fusion glycoprotein (F) have indicated that turkey rhinotracheitis virus (TRTV) is a pneumovirus, subfamily Pneumovirinae of the Paramyxoviridae family. As TRTV was known to generate polycistronic mRNAs, cDNA was generated from TRTV strain UK/3BV/85-infected Vero cell mRNAs using an oligonucleotide primer corresponding to a region of the F gene. Sequencing of four cDNAs revealed that the gene adjacent to the beginning (3' end) of the F gene was that for the matrix (M) protein, i.e., that TRTV had the partial gene order 3'-M-F-5'. This was unexpected as human respiratory syncytial (RS) virus, the type species of the genus Pneumovirus, has the partial gene order 3'-M-SH-G-F-5', where SH and G are the small hydrophobic protein and attachment glycoprotein, respectively. Instead TRTV resembled the Morbillivirus and Paramyxovirus genera of the Paramyxoviridae (subfamily Paramyxovirinae) which have the partial gene order 3'-M-F-5'. Two further oligonucleotides, one corresponding to a sequence near the end of the M gene and the other (oligo B) to a sequence near the beginning of the F gene, with their 5' ends spaced 300 nucleotides apart on the basis of the cDNA sequence, were used in a polymerase chain reaction (PCR) using genomic RNA as template. Only a PCR product of 0.3 kb was obtained. The same sized product was also obtained using these oligonucleotides and genomic RNA from three other TRTV strains (SA/91/78, UK/8544/85, and SA/2381/88) which had been grown in chicken tracheal organ cultures. In addition PCR was performed using genomic RNA from TRTV-3BV and SA/2381/88 with oligo B and another oligonucleotide near the 5' end of the gene upstream from M, spaced 1141 nucleotides apart on the basis of the sequence data. Only a 1.14-kb PCR product was obtained. Larger products would have been expected if another gene had been situated between M and F. The absence of such larger products, plus the demonstration that infected cells contained M-F dicistronic mRNAs, supported the conclusion that in the TRTV genome the M gene is adjacent to the F gene in the order 3'-M-F-5'. The 5' termini of the M and F mRNAs were confirmed by mRNA mapping. The TRTV M gene encoded a protein of 254 amino acids, very similar to that of RS virus (256 residues; 37% amino acid identity) but very different from that of the morbilliviruses and paramyxoviruses (approximately 350 residues).(ABSTRACT TRUNCATED AT 400 WORDS)
Yu
Q
Q
Institute for Animal Health, Houghton Laboratory, Huntingdon, Cambridgeshire, United Kingdom.
Davis
P J
PJ
Li
J
J
Cavanagh
D
D
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D002642:Chick Embryo; D003001:Cloning, Molecular; D004279:DNA, Viral; D005814:Genes, Viral; D008040:Genetic Linkage; D008969:Molecular Sequence Data; D010252:Paramyxoviridae / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012136:Respiratory Syncytial Viruses / Q000235:genetics*; D016415:Sequence Alignment; D014709:Vero Cells; D014763:Viral Matrix Proteins / Q000235:genetics*
D004279:DNA, Viral; D014763:Viral Matrix Proteins