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Plant Cell Advance Online Publication
Published on January 4, 2008; 10.1105/tpc.107.056606

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Received October 30, 2007
Returned for revision December 12, 2007
Accepted December 13, 2007

A Genomic Map of Viroid RNA Motifs Critical for Replication and Systemic Trafficking

Xuehua Zhong 1, Anthony J. Archual 1, Amy A. Amin 1, and Biao Ding 2*

1 Department of Plant Cellular and Molecular Biology, Plant Biotechnology Center, Ohio State University, Columbus, Ohio 43210
2 Department of Plant Cellular and Molecular Biology, Plant Biotechnology Center, Ohio State University, Columbus, Ohio 43210; Molecular, Cellular, and Developmental Biology Graduate Program, Ohio State University, Columbus, Ohio 43210; The RNA Group, Ohio State University, Columbus, Ohio 43210

* To whom correspondence should be addressed. E-mail: ding.35{at}osu.edu.

RNA replication and systemic trafficking play significant roles in developmental regulation and host–pathogen interactions. Viroids are the simplest noncoding eukaryotic RNA pathogens and genetic units that are capable of autonomous replication and systemic trafficking and offer excellent models to investigate the role of RNA structures in these processes. Like other RNAs, the predicted secondary structure of a viroid RNA contains many loops and bulges flanked by double-stranded helices, the biological functions of which are mostly unknown. Using Potato spindle tuber viroid infection of Nicotiana benthamiana as the experimental system, we tested the hypothesis that these loops/bulges are functional motifs that regulate replication in single cells or trafficking in a plant. Through a genome-wide mutational analysis, we identified multiple loops/bulges essential or important for each of these biological processes. Our results led to a genomic map of viroid RNA motifs that mediate single-cell replication and systemic trafficking, respectively. This map provides a framework to enable high-throughput studies on the tertiary structures and functional mechanisms of RNA motifs that regulate viroid replication and trafficking. Our model and approach should also be valuable for comprehensive investigations of the replication and trafficking motifs in other RNAs.






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