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Short Communication |
1 Division of Virology, Faculty of Biomedical and Life Sciences, University of Glasgow, Glasgow G11 5JR, UK
2 University of California San Francisco, Genentech Hall, San Francisco, CA 94143-2280, USA
Correspondence
David Evans
david.evans{at}vir.gla.ac.uk
Nucleotides in the terminal loop of the poliovirus 2C cis-acting replication element (2CCRE), a 61 nt structured RNA, function as the template for the addition of two uridylate (U) residues to the viral protein VPg. This uridylylation reaction leads to the formation of VPgpUpU, which is used by the viral RNA polymerase as a nucleotide–peptide primer for genome replication. Although VPg primes both positive- and negative-strand replication, the specific requirement for 2CCRE-mediated uridylylation for one or both events has not been demonstrated. We have used a cell-free in vitro translation and replication reaction to demonstrate that 2CCRE is not required for the initiation of the negative-sense strand, which is synthesized in the absence of 2CCRE-mediated VPgpUpU formation. We propose that the 3' poly(A) tail could serve as the template for the formation of a VPg–poly(U) primer that functions in the initiation of negative-sense strands.
Present address: School of Animal and Microbial Sciences, University of Reading, PO Box 228, Reading, UK.
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