Nucleic Acids Research, Vol 27, Issue 2 485-490, Copyright © 1999 by Oxford University Press
J Wang, JM Bakkers, JM Galama, HJ Bruins Slot, EV Pilipenko, VI Agol and WJ Melchers
The origin of replication ( oriR ) involved in the initiation of (-) strand
enterovirus RNA synthesis is a quasi-globular multi-domain RNA structure
which is maintained by a tertiary kissing interaction. The kissing
interaction is formed by base pairing of complementary sequences within the
predominant hairpin-loop structures of the enteroviral 3' untranslated
region. In this report, we have fully characterised the kissing
interaction. Site-directed mutations which affected the different base
pairs involved in the kissing interaction were generated in an infectious
coxsackie B3 virus cDNA clone. The kissing interaction appeared to consist
of 6 bp. Distortion of the interaction by mispairing of each of the base
pairs involved in this higher order RNA structure resulted in either
temperature sensitive or lethal phenotypes. The nucleotide constitution of
the base which gaps the major groove of the kissing domain was not relevant
for virus growth. The reciprocal exchange of the complete sequence involved
in the kissing resulted in a mutant virus with wild type virus growth
characteristics arguing that the base pair constitution is of less
importance for the initiation of (-) strand RNA synthesis than the
existence of the tertiary structure itself.
ARTICLES
Structural requirements of the higher order RNA kissing element in the enteroviral 3'UTR
University of Nijmegen, Department of Medical Microbiology, PO Box 9101, 6500 HB Nijmegen, The Netherlands.
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