Mutations within noncoding terminal sequences of model RNAs of Sendai virus: influence on reporter gene expression.
A reverse-genetics system employing the chloramphenicol acetyltransferase (CAT) reporter gene has been established previously for Sendai virus. We utilized PCR-directed mutagenesis to introduce nucleotide additions, deletions, and/or substitutions within terminal Sendai virus RNA sequences. The influence of these mutations on replication-transcription of these model Sendai-CAT RNAs was analyzed by assaying CAT activity. Results from these experiments indicate that (i) Sendai-CAT RNAs expressing wild-type levels of CAT activity conform to the Sendai virus rule of six, (ii) apparent exceptions to the rule of six exist in that the 5' terminus of the Sendai-CAT RNA is more tolerant than the 3' terminus of nucleotide additions or deletions, and (iii) the 3' leader region of Sendai-CAT RNA appears to be sensitive not only to mutagenesis (single-nucleotide addition or deletion) but also to changes in its total nucleotide length.
Harty
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Department of Microbiology, Mount Sinai School of Medicine, New York, New York 10029, USA.
Palese
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P
D001483:Base Sequence; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D015870:Gene Expression; D017930:Genes, Reporter; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D010222:Parainfluenza Virus 1, Human / Q000235:genetics*; D012367:RNA, Viral / Q000235:genetics*; D017384:Sequence Deletion
D012367:RNA, Viral; D015500:Chloramphenicol O-Acetyltransferase