Mutational analysis of the avian pneumovirus conserved transcriptional gene start sequence identifying critical residues.
Seven of the eight genes in the avian pneumovirus (APV) genome contain a conserved 9 nt transcriptional start sequence with the virus large (L) polymerase gene differing from the consensus at three positions. The sequence requirements of the APV transcriptional gene start sequence were investigated by generating a series of mutations in which each of the nine conserved bases was mutated to each of the other three possible nucleotides in a minigenome containing two reporter genes. The effect of each mutation was assessed by measuring the relative levels of expression from the altered and unaltered gene start sequences. Mutations at positions 2, 7 and 9 significantly reduced transcription levels while alterations to position 5 had little effect. The L gene start sequence directed transcription at levels approximately 50 % below that of the consensus gene start sequence. These data suggest that there are common features in pneumovirus transcriptional control sequences.
Edworthy
Nicole L
NL
Department of Biological Sciences, University of Warwick, Coventry CV4 7AL, UK.
Easton
Andrew J
AJ
Department of Biological Sciences, University of Warwick, Coventry CV4 7AL, UK.
D015500:Chloramphenicol O-Acetyltransferase / Q000032:analysis; D017124:Conserved Sequence; D004252:DNA Mutational Analysis; D015870:Gene Expression; D017930:Genes, Reporter; D015751:Genes, pol; D008156:Luciferases / Q000032:analysis; D029121:Metapneumovirus / Q000235:genetics*; D016297:Mutagenesis, Site-Directed; D012333:RNA, Messenger / Q000032:analysis; D012367:RNA, Viral / Q000032:analysis; D024363:Transcription Initiation Site; D014158:Transcription, Genetic
D012333:RNA, Messenger; D012367:RNA, Viral; D008156:Luciferases; D015500:Chloramphenicol O-Acetyltransferase