1887

Abstract

Avocado sunblotch viroid (ASBVd), the type member of the family , replicates and accumulates in chloroplasts. Whether this minimal non-protein-coding circular RNA of 246–250 nt exists as a free nucleic acid or closely associated with host proteins remains unknown. To tackle this issue, the secondary structures of the monomeric circular (+) and (−) strands of ASBVd have been examined by searching those of minimal free energy, and at single-nucleotide resolution by selective 2′-hydroxyl acylation analysed by primer extension (SHAPE). Both approaches resulted in predominant rod-like secondary structures without tertiary interactions, with the (+) RNA being more compact than its (−) counterpart as revealed by non-denaturing polyacryamide gel electrophoresis. Moreover, SHAPE showed that the ASBVd (+) form accumulates in avocado leaves as a free RNA adopting a similar rod-shaped conformation unprotected by tightly bound host proteins. Hence, the ASBVd (+) RNA behaves like the previously studied (+) RNA of potato spindle tuber viroid, the type member of nuclear viroids (family ), indicating that two different viroids replicating and accumulating in distinct subcellular compartments, have converged into a common structural solution. Circularity and compact secondary structures confer to these RNAs, and probably to all viroids, the intrinsic stability needed to survive in their natural habitats. However, SHAPE has not revealed the (possibly transient or loose) interactions of the ASBVd (+) RNA with two host proteins observed previously by UV irradiation of infected avocado leaves.

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2017-07-01
2020-01-19
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