Nucleic Acids Research, Vol 26, Issue 2 420-426, Copyright © 1998 by Oxford University Press
A Ashoub, W Rohde and D Prufer
The genetic information of potato leafroll virus (PLRV), a typical member
of the subgroup 2 luteoviruses, is contained in a single- stranded (+)
sense RNA of approximately 5.9 kb. A single subgenomic RNA (sgRNA1) of
approximately 2.3 kb has been characterized as the mRNA for the 3'
clustered viral open reading frames ORF3, ORF3/5 and ORF4. Here we
demonstrate by Northern blot analyses of polysomal RNAs from PLRV- infected
Solanum tuberosum and Physalis floridana plants that, as with luteoviruses
belonging to subgroup 1, in planta synthesis of a second 0.8 kb subgenomic
RNA (sgRNA2) increases the complexity of subgroup 2 luteoviral genomes
significantly. PLRV-specific hybridization probes as well as primer
extension experiments map sgRNA2 to the 3'-end of the PLRV RNA genome
(positions 5190-5987). Similarly, for the closely related cucurbit
aphid-borne yellows virus (CABYV) a sgRNA2 of similar size and position
(positions 4888-5669) was identified. PLRV sgRNA2 may code for two viral
proteins of 7.1 (ORF6) and 14 kDa (ORF7) respectively, while the CABYV
proteins are 8.7 (ORF6) and 8.3 kDa (ORF7) in size, with PLRV ORF7
displaying nucleic acid binding activity. In vivo experiments by transient
expression of chimeric GUS fusions in potato protoplasts demonstrated that
sgRNA2 functions as a bicistronic mRNA with high expression of ORF6 and low
translational efficiency for synthesis of ORF7.
ARTICLES
In planta transcription of a second subgenomic RNA increases the complexity of the subgroup 2 luteovirus genome
Max-Planck-Institut fur Zuchtungsforschung, Carl-von-Linne-Weg 10, D- 50829 Koln, Germany.
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