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J. Virol., Aug 1997, 5990-5996, Vol 71, No. 8
BA Deiman, RM Kortlever and CW Pleij
The tRNA-like structure at the 3' end of turnip yellow mosaic virus (TYMV)
RNA was studied in order to determine the role of this structure in the
initiation of minus-strand synthesis in vitro. Deletions in the 5'-to-3'
direction up to the pseudoknot structure did not result in a decrease of
transcription efficiency. However, transcription efficiency was reduced
twofold when a fragment of 21 nucleotides, comprising the 3'-terminal
hairpin, was used as a template. tRNA(Phe) from yeast, Escherichia coli 5S
rRNA, and the 3'-terminal 208 nucleotides of alfalfa mosaic virus RNA 3
could not be transcribed by the RNA- dependent RNA polymerase (RdRp) of
TYMV. Various mutations in the sequences of loop regions L1 and L2 or of
stem region S1 of the pseudoknot were tested to further investigate the
importance of the pseudoknot structure. The results were compared with
those obtained in an earlier study on aminoacylation with the same mutants
(R. M. W. Mans, M. H. van Steeg, P. W. G. Verlaan, C. W. A. Pleij, and L.
Bosch, J. Mol. Biol. 223:221-232; 1992). Mutants which still harbor a
stable pseudoknot, as proven by probing its structure, have a transcription
efficiency very close to that of the wild-type virus. Disruption of the
pseudoknot structure, however, gives rise to a drop in transcription
efficiency to about 50%. No indications of base-specific interactions
between L1, L2, or S1 of the pseudoknot and the RdRp were found.
Copyright © 1997, American Society for Microbiology
The role of the pseudoknot at the 3' end of turnip yellow mosaic virus RNA in minus-strand synthesis by the viral RNA-dependent RNA polymerase
Leiden Institute of Chemistry, Gorlaeus Laboratories, The Netherlands.
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