Cy. Huang et al., Sequences at the 3 ' untranslated region of bamboo mosaic potexvirus RNA interact with the viral RNA-dependent RNA polymerase, J VIROLOGY, 75(6), 2001, pp. 2818-2824
The 3' untranslated region (UTR) of bamboo mosaic potexvirus (BaMV) genomic
RNA was found to fold into a series of stem-loop structures including a ps
eudoknot structure. These structures were demonstrated to be important for
viral RNA replication and were believed to be recognized by the replicase (
C.-P. Cheng and C.-H. Tsai, J. Mel. Biol. 288:555-565, 1999). Electrophoret
ic mobility shift and competition assays have now been used to demonstrate
that the Escherichia coli-expressed RNA-dependent RNA polymerase domain (De
lta 893) derived from BaMV open reading frame 1 could specifically bind to
the 3' UTR of BaMV RNA. No competition was observed when bovine liver tRNAs
or poly(I)(C) double-stranded homopolymers,were used as competitors, and t
he cucumber mosaic virus 3' UTR was a less efficient competitor. Competitio
n analysis with different regions of the BaMV 3' UTR shelved that Delta 893
binds to at least two independent RNA binding sites, stem-loop D and the p
oly(A) tail. Footprinting analysis revealed that Delta 893 could protect th
e sequences at Loop D containing the potexviral conserved hexamer motif and
part of the stem of domain D from chemical cleavage.