Role of the nucleocapsid protein in regulating vesicular stomatitis virus RNA synthesis

Abstract

We describe experiments with two monoclonal antibodies to the vesicular stomatitis virus (VSV) nucleocapsid protein N with strikingly different characteristics. Antibody 1 binds to nucleocapsids and probably the pool of free (unbound) N protein; it inhibits transcription in vitro, and when microinjected into cells, protects the cells against VSV. Antibody 2 binds poorly to nucleocapsids, does not inhibit transcription, but when microinjected into cells, binds selectively to the free N and delays the appearance of progeny virus. We have confirmed these results by analyzing the effect of these antibodies on in vitro genomic RNA synthesis. The results of both the in vivo and in vitro experiments show that the replication of the VSV genome is controlled by the availability of the nucleocapsid protein, even when the polymerase has access to the host factors and multiple phosphorylated forms of the NS protein thought to be involved in genomic RNA synthesis.