JBC, Vol. 252, Issue 3, 872-877, Feb, 1977
High molecular weight virion-associated RNA of vaccinia. A possible precursor to 8 to 12 S mRNA
E. Paoletti
The high molecular weight virion-associated RNA synthesized by vaccinia in
vitro can be cleaved into smaller components, some of which are extruded
from the virus as 8 to 12 S RNA. The high molecular weight
virion-associated RNA fails to bind appreciably (5%) to poly(U) filters
indicating that it is not polyadenylated. Its cleavage products will,
however, bind to poly(U) (40 to 50%) after processing in the presence of
ATP. The high molecular weight virion-associated RNA is methylated by the
virus, and purified unmethylated RNA can be methylated by
detergent-solubilized extracts of vaccinia virus cores. In the presence of
GTP, methylation is stimulated 3-fold. The level of methylation of purified
unmethylated high molecular weight RNA achieved by soluble core extracts is
approximately 80% of the level of methylation achieved with purified
unmethylated 8 to 12 S viral RNA, suggesting that more residues than the
primary 5' termini became methylated. Approximately 85% of the methylated
RNA binds to poly(U) when purified high molecular weight RNA is processed
with soluble core extracts in the presence of
S-adenosyl[methyl-3H]methionine, GTP, and ATP, conditions which also cleave
the RNA. Nucleic acid hybridization-competition studies indicate that
virion-extruded 8 to 12 S mRNA contains sequences found in the high
molecular weight virion-associated RNA.
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