Characterization of vesicular stomatitis virus mRNA species synthesized in vitro

Abstract

The smallest size class of mRNA (12S) synthesized in vitro by the virionassociated RNA polymerase of vesicular stomatitis virus contains two mRNA species of similar molecular weight that code for the viral M and NS proteins. The resolution of these mRNA species was achieved by converting them to duplexes by annealing with the genome RNA, followed by RNase T2 treatment and separation in a polyacrylamide gel. Using this separation technique, the mRNA's were identified by comparing the relative resistance of their syntheses to UV irradiation of the virus. The molecular weights of these two mRNA species calculated as duplex RNAs were smaller than expected. The possible reasons for this discrepancy are discussed.

Transcription and replication of rhabdoviruses.

“…(i) All mRNA species are initiated with an identical 5'-terminal sequence, G(5')ppp(5')AACCAG (182), whereas the 3'-terminal sequence of the genome RNA is 3' UGC, which is not complementary to any of the mRNAs (17). (ii) The synthesis of mRNA species, both in vitro and in vivo, is nonequimolar; i.e., the synthesis of N mRNA is in the highest molar amount, followed by NS, M, G, and L, in that order (180,181,231). (iii) The unique 47-base-long leader RNA is the first product to be synthesized in vitro (52).…”

Assignment of the large oligonucleotides of vesicular stomatitis virus to the N, NS, M, G, and L genes and oligonucleotide gene ordering within the L gene

“…1B. Five major bands of labeled material were observed, corresponding (in order of increasing electrophoretic mobility) to L, G, N, NS, and M duplexes (11,15). Faint bands of radioactivity were also evident, possibly representing nicked duplexes or duplexes derived from multistranded hybrids (Fig.…”

“…The New Mexico variant of VSV Indiana does not have 11 oligonucleotides that are present in the prototype strain (no. 7,10,15,16,24,25,49,57,58,63, and 65). Of these oligonucleotides, seven have been assigned to the L gene (no.…”

Proteins of vesicular stomatitis virus. V. Identification of a precursor to the phosphoprotein of Piry virus

“…The possible precursor-product relationship of the two proteins was demonstrated by pulsechase experiments. Piry virus infected celLs were pulsed with [3S]methionine followed by incu- the conversion of NSi to NS, occurs with a halflife of approximately 20 min under the conditions employed.…”

“…An extract of labeled uninfected cells is presented in the central well (CNT). on September 28, 2020 by guest http://jvi.asm.org/ Downloaded from greater than the coding capacity of the RNA for this protein (7,20), makes any attempt to correlate electrophoretic mobility and molecular weight for this protein of doubtful significance. Since shape factors and/or degree of binding of em.~~~~~~O and absence of cycloheximide.…”