Nucleic Acids Research, Vol 24, Issue 11 2133-2142, Copyright © 1996 by Oxford University Press
S Todd and BL Semler
The specific recognition of genomic positive strand RNAS as templates for
the synthesis of intermediate negative strands by the picornavirus
replication machinery is presumably mediated by cis-acting sequences within
the genomic RNA 3' non-coding region (NCR). A structure- infectivity
analysis was conducted on the 44 nt human rhinovirus 14 (HRV14) 3' NCR to
identify the primary sequence and/or secondary structure determinants
required for viral replication. Using biochemical RNA secondary structure
probing techniques, we have demonstrated the existence of a single
stem-loop structure contained entirely within the 3' NCR, which appears to
be phylogenetically conserved within the rhinovirus genus. We also report
the in vivo analysis of a number of 3' NCR deletion mutations engineered
into infectious cDNA clones which were designed to disrupt the stem-loop
secondary structure to varying degrees. Large deletions (up to 37 nt)
resulted in defective growth phenotypes, although they were not lethal. We
propose that the absolute requirements for initiation of negative strand
synthesis are less stringent than previously postulated, even though
defined RNA secondary structure determinants may have evolved to facilitate
and/or regulate the process of viral RNA replication.
ARTICLES
Structure-infectivity analysis of the human rhinovirus genomic RNA 3' non-coding region
Department of Microbiology and Molecular Genetics, College of Medicine, University of California, Irvine 92717, USA.
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