Sendai virus contains overlapping genes expressed from a single mRNA

SUMMARYA cDNA library was prepared from poly(A) + RNA extracted from respiratory syncytial (RS) virus-infected HEp-2 cells. A recombinant plasmid, pRSP68, encoding the RS virus phosphoprotein gene was identified by translation in vitro of hybridselected mRNA. The cloned cDNA insert of 1 kb was sequenced and a polypeptide of 241 amino acids with a molecular weight of 27166 was deduced from the sequence. The protein was relatively rich in polar amino acids and devoid of both cysteine and tryptophan. A second short open reading frame with a coding potential of 65 amino acids was identified and overlapped the 3' terminus of the phosphoprotein gene by 34 bases.Respiratory syncytial (RS) virus is the major pathogen causing severe lower respiratory tract infection in infants (Parrott et al., 1973). The virus is a pleomorphic, unsegmented, negativestranded RNA virus similar to the paramyxoviruses but placed in a separate genus, Pneumovirus, on the basis of its morphology and the lack of detectable neuraminidase and haemagglutinin activities. More recently, study of the genetic organization of RS virus has highlighted the differences between it and the typical paramyxoviruses . The RS viral genome is a single negative strand of RNA with a molecular weight of around 5 × 106, and is transcribed into 10 discrete polyadenylated mRNAs coding for 10 unique viral proteins . The gene order has been determined by transcriptional mapping using u.v. inactivation kinetics (Dickens et al., 1984). These studies provided evidence of a single promoter site for RS virus transcription and the gene order:

mentioning

confidence: 76%

Nucleotide Sequence of the Respiratory Syncytial Virus Phosphoprotein Gene

Lambden

1985

“…I for the Indiana serotype, Mudd-Summers strain) which could encode a small (65 or 67 amino acid) basic protein. The phosphoprotein genes of two other negative-strand viruses, measles and Sendai viruses, also contain other ORFs; moreover, these internal ORFs are translated efficiently in vitro and during viral infection (Bellini et al, 1985;Giorgi et al, 1983;Curran et al, 1986). Although the precise function of this class of protein (named C protein) is unknown, the co-localization of the measles C protein with the nucleocapsid protein suggests a role in replication or transcription (Bellini et al, 1985).…”

Section: Sequence Of the Cloned Ns And Intergenic Regionsmentioning

confidence: 99%

Cloning and Expression of a Viral Phosphoprotein: Structure Suggests Vesicular Stomatitis Virus NS May Function by Mimicking an RNA Template

Hudson¹,

Condra²,

Lazzarini³

1986

SUMMARYThe phosphoprotein (NS) gene from the Indiana serotype of vesicular stomatitis virus (VSV; Mudd-Summers strain) was cloned and sequenced. The NS gene encodes a protein of 265 amino acids which was expressed from a simian virus 40 vector in COS cells. The post-translational modification characteristic of viral NS, the extensive phosphorylation of a cluster of serine and threonine residues, was also evident in recombinant NS protein. The NS gene displays a property common to the phosphoprotein genes of negative-strand RNA viruses: the phosphoprotein mRNA has a second open reading frame (ORF) which could encode a small (7500 mol. wt.) protein. Both measles virus and Sendai virus employ the second ORF of their phosphoprotein gene, and the resultant proteins have an amino acid composition similar to that predicted for the VSV ORF. Comparison of phosphoproteins from different VSV strains revealed two conserved domains that we propose are critical for the function of NS in transcription and replication.

“…The previous studies are not fully in accord, however, as to whether the non-structural proteins are related to the P protein or not (Herder & Compans, 1982;Merz et al, 1983;Simpson et al, 1984). It is known that many paramyxoviruses such as Sendal virus (Giorgi et aL, 1983 ;Shioda et al, 1983), human parainfluenza virus 3 (PF-3) (Galinski et al, 1986;Luk et al, 1986;Spriggs & Collins, 1986), measles virus (Bellini et al, 1985) and canine distemper virus (Barrett et al, 1985) utilize different reading frames present in the P protein mRNA to code for the non-structural C proteins. By sequencing cloned cDNA of the P gene region, we attempted to determine whether a bicistronic coding strategy is also used in the mumps virus P gene.…”

Section: Introductionmentioning

confidence: 99%

Molecular Cloning and Sequence Analysis of the Mumps Virus Gene Encoding the P Protein: Mumps Virus P Gene Is Monocistronic

Takeuchi

Hishiyama²,

Yamada³

et al. 1988

SUMMARYThe nucleotide sequence of the P (phosphoprotein) gene of two strains of mumps virus has been determined from overlapping cDNA clones. The P gene contained a single open reading frame coding for a protein of 391 amino acids with a calculated Mr of 41587, in good agreement with the value (40K to 45K) estimated from electrophoretic mobility on SDS-polyacrylamide gels. No open reading frame analogous to the C gene of other paramyxoviruses existed in the mumps virus P gene region. Comparison of the amino acid sequence of the mumps virus P protein with that of Newcastle disease virus showed a limited sequence homology.