Identification of a cis -Acting Replication Element within the Poliovirus Coding Region

• DOI: 10.1128/jvi.74.10.4590-4600.2000
• Publication Date: 2002-07-27T10:06:23Z
• Authors: Ian Goodfellow; Yasmin Chaudhry; Andrew Richardson; Janet Meredith; Jeffrey W. Almond; Wendy Barclay; David J. Evans
• Keywords: Subgenomic mRNA; Picornavirus; Replicon

ABSTRACT The replication of poliovirus, a positive-stranded RNA virus, requires translation the infecting genome followed by virus-encoded VPg and 3D polymerase-primed synthesis negative-stranded template. sequences involved in latter process are poorly defined. Since many picornavirus form structures, we searched genome, other than untranslated regions, for predicted local secondary structural elements identified 61-nucleotide (nt) stem-loop region encoding 2C protein. Covariance analysis suggested structure was well conserved Enterovirus genus Picornaviridae . Site-directed mutagenesis, disrupting without affecting product, destroyed viability that required positive sense function. Recovery revertant viruses integrity critical function, demonstrated nonviable mutants did not synthesize negative strands. Our conclusion, this constitutes novel poliovirus cis -acting element (CRE), is supported demonstration subgenomic replicons bearing lethal mutations native can be restored to competence addition second copy 61-nt wild-type sequence at another location within genome. This CRE functionally resembles an rhinovirus type 14 (K. L. McKnight S. M. Lemon, 4:1569–1584, 1998) cardioviruses (P. E. Lobert, N. Escriou, J. Ruelle, T. Michiels, Proc. Natl. Acad. Sci. USA 96:11560–11565, 1999) but differs sequence, structure, location. functional role evolutionary significance CREs positive-sense discussed.