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Journal of Virology, December 1999, p. 9952-9958, Vol. 73, No. 12
School of Animal and Microbial Sciences,
Received 15 March 1999/Accepted 31 August 1999
We showed previously that a human rhinovirus 14 (HRV14) 3'
untranslated region (3' UTR) on a poliovirus genome was able to replicate with nearly wild-type kinetics (J. B. Rohll, D. H. Moon, D. J. Evans, and J. W. Almond, J. Virol
69:7835-7844, 1995). This enabled the HRV14 single 3' UTR stem-loop
structure to be studied in combination with a sensitive reporter
system, poliovirus FLC/REP, in which the capsid coding region is
replaced by an in-frame chloramphemicol acetyltransferase (CAT) gene.
Using such a construct, we identified a mutant (designated mut4), in
which the structure and stability of the stem were predicted to be
maintained, that replicated very poorly as determined by its level of
CAT activity. The effect of this mutant 3' UTR on replication has been
further investigated by transferring it onto the full-length cDNAs of
both poliovirus type 3 (PV3) and HRV14. Virus was recovered with a
parental plaque phenotype at a low frequency, indicating the
acquisition of compensating changes, which sequence analysis revealed
were, in both poliovirus- and rhinovirus-derived viruses, located in
the active-site cleft of 3D polymerase and involved the substitution of
Asn18 for Tyr. These results provide further evidence of a specific
interaction between the 3' UTR of picornaviruses and the viral
polymerase and also indicate similar interactions of the 3' UTR of
rhinovirus with both poliovirus and rhinovirus polymerases.
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Similar Interactions of the Poliovirus and
Rhinovirus 3D Polymerases with the 3' Untranslated Region of
Rhinovirus 14
and
*
Corresponding author. Mailing address: Institute of
Virology, University of Glasgow, Church St., Glasgow, G11 5JR, United Kingdom. Phone and fax: 44 (0)141 330 6249. E-mail:
David.Evans{at}vir.gla.ac.uk.
Present address: Pasteur Mérieux Connaught, 69280 Marcy-L'Etoile, France.
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