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Nucleic Acids Research, Vol 25, Issue 23 4723-4729, Copyright © 1997 by Oxford University Press

ARTICLES

Conformation of the 3'-end of beet necrotic yellow vein benevirus RNA 3 analysed by chemical and enzymatic probing and mutagenesis

E Lauber, H Guilley, K Richards, G Jonard and D Gilmer
Institut de Biologie Moleculaire des Plantes du CNRS et de l'Universite Louis Pasteur, 12 Rue du General Zimmer, 67084 Strasbourg Cedex, France.

Secondary structure-sensitive chemical and enzymatic probes have been used to produce a model for the folding of the last 68 residues of the 3'-non-coding region of beet necrotic yellow vein benevirus RNA 3. The structure consists of two stem-loops separated by a single-stranded region. RNA 3-derived transcripts were produced containing mutations which either disrupted base pairing in the helices or maintained the helices but with alterations in the base pairing scheme. Other mutants contained substitutions in single-stranded regions (loops or bulged sequences). With a few exceptions all three types of mutation abolished RNA 3 replication in vivo, suggesting that both secondary structure and specific sequences are required for efficient recognition of the 3'- terminal region of RNA 3 by viral RNA-dependent RNA polymerase.
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