RNase T(1)
oligonucleotide fingerprint analyses of three
vesicular stomatitis virus Indiana serotype small defective interfering (DI) particle
RNA species indicate that they only have
oligonucleotides derived from the 5' region of the viral genome. These studies also indicate that these three DI RNAs have partial L gene sequences as well as two 5' viral
oligonucleotides (59 and 70) that are not transcribed into L (or other)
mRNA species (J. P. Clewley and D. H. L. Bishop, J. Virol. 30:116-123, 1979). Analyses of the large DI
RNA (LT DI) reveal a different origin. The LT DI
RNA has
oligonucleotides derived from both the 3' end of the genome (including all the large
oligonucleotides identified for N, NS, M, and G genes), in addition to at least one of the 5'-proximal L gene
oligonucleotides (47), as well as all seven
oligonucleotides (3, 38, 42, 43, 44B, 59, and 70) that are not protected from nuclease digestion after the formation of
mRNA-
viral RNA duplexes (Clewley and Bishop). It appears therefore that the genesis of LT
RNA involves a deletion of internal L gene sequences from the
viral RNA.
Oligonucleotide sequence analyses have been undertaken on several of the
vesicular stomatitis viral RNA oligonucleotides, including all seven (3, 38, 42, 43, 44B, 59, and 70) that are not transcribed into
mRNA. The analyses confirm that
oligonucleotides 59 [3'...GAACACCAAAAAUAAAAAAUA(G)...5'] and 70 [3'...GACCAAAACACCA(G)...5'] are at the 5'-end region of the viral genome.
Oligonucleotide 38 [3'...GAAAUUCAUACUUUUUU(U)(G)...5'] may represent the termination signal for L
mRNA synthesis (R. A. Lazzarini, personal communication).
Oligonucleotide 43 [3'...GUAUACUUUUUUU(G)...5'] corresponds to the sequence shown to be the N gene
mRNA polyadenylation signal (D. J. McGeoch, Cell 17:673-681, 1979). The other three
oligonucleotides share a common feature with
oligonucleotides 43 and 38, viz., a stretch of 6 or 7 U residues preceded by an AUAC sequence. Thus the sequence of
oligonucleotide 3 is 3'...GAAUUAAUAUAAAAUUAAAAAUUAAAAAUACUUUUUU(U)(G)...5', whereas that of
oligonucleotide 42 is 3'...GAUACUUUUUUUCAU(U)(G)...5', and that of
oligonucleotide 44B is 3'...G(U)AUACUUUUUU(G)...5'. These sequence analyses suggest a common polyadenylation signal for the synthesis of all
vesicular stomatitis virus
mRNA species, i.e., the sequence (3')...AUACUUUUUU(U)...(5').