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Abstract

The 3′ untranslated region (UTR) of turnip crinkle virus (TCV) RNA is 253 nt long (nt 3798–4050) with a 27 nt hairpin structure near its 3′ terminus. In this study, the roles of the 3′ UTR in virus accumulation were investigated in protoplasts of L. and (L.) Heynh. Our results showed that, in protoplasts, the minimal 3′ UTR essential for TCV accumulation extends from nt 3922 to 4050, but that maintenance of virus accumulation at wild-type (wt) levels requires the full-length 3′ UTR. However, in protoplasts, only 33 nt (nt 4018–4050) at the 3′ extremity of the UTR is required for wt levels of accumulation, whereas other parts of the 3′ UTR are dispensable. The 27 nt hairpin within the 33 nt region is essential for virus accumulation in both and protoplasts. However, transposition of nucleotides in base pairs within the upper or lower stems has no effect on virus accumulation in either or protoplasts, and alterations of the loop sequence also fail to affect replication. Disruption of the upper or lower stems and deletion of the loop sequence reduce viral accumulation in protoplasts, but abolish virus accumulation in protoplasts completely. These results indicate that strict conservation of the hairpin structure is more important for replication in than in protoplasts. In conclusion, both the 3′ UTR primary sequence and the 3′-terminal hairpin structure influence TCV accumulation in a host-dependent manner.

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2007-02-01
2020-01-24
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