Ribonucleic Acid Synthesis of Vesicular Stomatitis Virus, II. An RNA Polymerase in the Virion

Abstract

Abstract. The virions of vesicular stomatitis virus contain an enzyme that catalyzes the incorporation of ribonucleotides into RNA. The product of the reaction is mainly RNA complementary in base sequence to that of vesicular stomatitis virus RNA.Introduction. Two considerations led us to search for an RNA polymerase in the virion of vesicular stomatitis virus, a single-stranded RNA virus. First, the lack of infectivity of vesicular stomatitis virus RNA,I which contrasts with the readily demonstrable infectivity of RNA from arboviruses2 or picornaviruses,3 could be explained by an obligate requirement for a virion polymerase in order to initiate a vesicular stomatitis virus infection. Second, the recent finding that the messenger RNA's that synthesize vesicular stomatitis virus proteins are complementary to the nucleotide sequence of the virion RNA4 led us to wonder how the initial stages of the infection proceed: if the virion RNA is not a messenger, then it must be able to serve as template for messenger RNA synthesis using either a polymerase found in the host cell or one that is a part of the virion.Two precedents exist for the association of an RNA polymerase with the virion of an animal virus. One is the finding of a DNA-dependent RNA polymerase in vaccinia virions.5 The second is the finding of an RNA-dependent RNA polymerase in the virions of reovirus6 and other double-stranded RNA viruses.7Materials and Methods. Purified B particles8 of vesicular stomatitis virus, the

Characterization of rabies virus nucleocapsids and recombinant nucleocapsid-like structures.

“…In most cases, the viral nucleocapsids are functional in vitro, either by just opening up the virus by detergent and adding NTPs (plus a primer for influenza virus) or after purification of the nucleocapsids (Baltimore et al, 1970 ;Leppert et al, 1979 ;Vidal & Kolakofsky, 1989 ;Honda et al, 1986 ;Ishihama et al, 1986). For VSV and Sendai virus it has been possible to separately purify the nucleoprotein-RNA complexes and L plus P (either from virus or from recombinant sources) and recover activity after reconstitution in vitro (Emerson & Wagner, 1972 ;Emerson & Yu, 1975 ;De & Banerjee, 1984 ;Masters & Banerjee, 1986 ;Helfman & Perrault, 1989 ;Emerson, 1982Emerson, , 1987Canter et al, 1993 ;Horikami et al, 1992 ;Curran et al, 1992).…”

“…Since the infected cells do not have a suitable enzyme for this activity, all viruses in this group carry their own RNA-dependent RNA polymerase (Baltimore et al, 1970). The genome of the negative-strand RNA viruses, whether segmented or not, is encapsidated by a virus-specific nucleoprotein (N).…”

Genetic manipulation of non-segmented negative-strand RNA viruses

“…1). With regard to their expression strategy, it is not surprising that isolated RNA from negative-strand RNA viruses is not infectious, a criterion which was originally used to distinguish positive-and negative-strand RNA viruses (Baltimore et al, 1970). Proteins are not expressed from the genomic RNA due to its negative polarity, or from the complementary positive strand due to the modular organization of the RNA, which does not allow translation of (all) virus proteins by the cellular machinery.…”

Vesicular stomatitis virus deficient in protein kinase is infectious

“…The RNA extracted from such virions is not infectious [2]. To exhibit infectivity, the virion RNA (negative-strand RNA) must be transcribed by the virion-associated transcriptase into a complementary RNA (positive-strand RNA) that serves as a messenger RNA for the synthesis of virusspecific proteins as well as a template for the production of virion RNA [ 1,3]. The transcriptase which has been purified was found to consist of two of the five viral structural polypeptides, the L and NS polypeptides [4].…”