The P protein and the nonstructural 38K and 29K proteins of newcastle disease virus are derived from the same open reading frame

The nucleotide sequence of cloned cDNA copies of the mRNA encoding the Newcastle disease virus (NDV), strain AV, phosphoprotein (P) was determined. The sequence of 1443 nucleotides contains one long open reading frame which could encode a protein with a molecular weight of 42,126, and two smaller open reading frames which could encode proteins with molecular weights of 11,178 and 13,935. Full-length cDNA clones were constructed in an SP6 vector, mRNA was transcribed in a cell-free system using the SP6 polymerase, and the mRNA was translated in a wheat germ cell-free extract. The P mRNA directed the synthesis of, primarily, four products. One, with a molecular weight of 53,000 Da, comigrated with authentic P protein made in infected cells and was precipitable with antisera with specificity for the NDV P protein. The other products of the cell-free reaction had molecular weights of 38,000, 29,000 and 12,000. The 29,000- and the 38,000-Da polypeptides were also precipitable with anti-P protein antibody. Using truncated cDNA clones, evidence is presented that the 38,000- and 29,000-Da proteins are derived from initiation at AUG triplets in the same reading frame as the P protein. Infected cells also contain these polypeptides which may be analogous to C proteins of other paramyxoviruses. Thus the NDV P protein mRNA is different than most other paramyxovirus P protein mRNAs which are translated in two different reading frames to yield the P and C proteins.