PREPARATION AND ANALYSIS OF THE NUCLEOCAPSID PROTEINS OF VESICULAR STOMATITIS-VIRUS AND SENDAI VIRUS, AND ANALYSIS OF THE SENDAI VIRUS LEADER-NP GENE REGION

A procedure is presented for isolating the nucleocapsid proteins, N and NP from vesicular stomatitis virus and Sendai virus, respectively, in soluble form. These proteins were suitable for the determination of their blocked amino-terminal peptide sequences by GLC/mass spectrometry at the low nanomole level. The N protein prepared by this procedure was previously shown to retain some of its expected biological activity. The sequence of 626 nucleotides from the 3'' end of the Sendai virus genome, which includes the first 1/3 of the NP gene, was determined. Using this information, primer extension studies on intracellular Sendai virus mRNA allowed the determination of the structure of the leader-NP intervening sequence and the 5'' end of the NP mRNA. Comparison of the amino termini of the nucleocapsid proteins with their respective mRNA sequences revealed that these proteins are similarly processed in vivo. [Hamster kidney BHK21 cells were used in this study.].