The mutation responsible for the temperature-sensitive (ts) phenotype of mutant N19 (complementation group E) of respiratory syncytial virus has been located to the P protein gene. Viral protein synthesis was completely restricted at 39 °C, and the N19 P protein did not react with an anti-P monoclonal antibody (MAb) (3–5) at 33 °C. Reversion of temperature sensitivity restored reactivity with MAb 3–5. Nucleotide sequence determination and expression of cDNA clones of P mRNA derived from wild-type, N19 and non-ts revertant-infected cells, revealed that temperature sensitivity and loss of reactivity with MAb 3–5 were consequences of a Gly → Ser amino acid change at position 172. A low polypeptide, which represented the C-terminal 93 amino acids of the P protein, was produced by internal initiation in the P open reading frame during translation, and a similar product was detected transiently .


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