Synthesis of leader RNA and editing of the P mRNA during transcription by purified measles virus.
A transcription system with detergent-disrupted purified measles virus was developed. Synthesis of authentic, full-length measles virus N, P, M, and F mRNAs by purified virus occurred as identified by dot-blot hybridization analysis of individual measles virus clones and gel electrophoresis. The relative abundance of the first five viral mRNAs synthesized in vitro decreased significantly with their distance from the 3' end. The addition of the soluble protein fraction from uninfected A549 cells stimulated overall viral RNA synthesis but did not alter the relative abundance of each of the mRNAs. Measles virus synthesized in vitro a leader RNA of approximately 55 nucleotides in length, suggesting that like other negative-strand viruses, transcription initiated only at the 3' end of the genome RNA. Purified measles virus also catalyzed RNA editing during the synthesis of the P mRNA as shown by modified primer extension analysis of the mRNA products and by translation of the modified RNA into the V protein in rabbit reticulocyte lysates. These data suggested that the RNA editing activity was virus encoded.
Horikami
S M
SM
Department of Immunology and Medical Microbiology, University of Florida School of Medicine, Gainesville 32610.
Moyer
S A
SA
D000818:Animals; D001483:Base Sequence; D003553:Cystine / Q000378:metabolism; D008459:Measles virus / Q000235:genetics* / Q000254:growth & development / Q000302:isolation & purification; D008715:Methionine / Q000378:metabolism; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics* / Q000302:isolation & purification; D012367:RNA, Viral / Q000235:genetics* / Q000302:isolation & purification; D014158:Transcription, Genetic; D014709:Vero Cells; D014764:Viral Proteins / Q000096:biosynthesis / Q000235:genetics
D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D012367:RNA, Viral; D014764:Viral Proteins; D003553:Cystine; D008715:Methionine