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Journal of Virology, April 1999, p. 2703-2709, Vol. 73, No. 4
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Sufficient Length of a Poly(A) Tail for the Formation of a Potential Pseudoknot Is Required for Efficient Replication of Bamboo Mosaic Potexvirus RNA

Ching-Hsiu Tsai,1,* Chi-Ping Cheng,1 Chih-Weng Peng,1 Biing-Yuan Lin,2 Na-Sheng Lin,2 and Yau-Heiu Hsu1

Graduate Institute of Agricultural Biotechnology, National Chung Hsing University, Taichung 402,1 and Institute of Botany, Academia Sinica, Nankang, Taipei 115,2 Taiwan

Received 11 September 1998/Accepted 4 December 1998

RNAs transcribed from a full-length infectious cDNA clone of the bamboo mosaic potexvirus (strain O) genome, pBaMV-O, were infectious to Nicotiana benthamiana plants. Mutant genomes in which the poly(A) tail is absent or replaced by a 3' tRNA-like structure from turnip yellow mosaic virus RNA failed to amplify detectably in N. benthamiana protoplasts. No amplification was detected in protoplasts inoculated with transcripts containing 4, 7, or 10 adenylate residues at the 3' end, whereas transcript inocula with 15 adenylate residues resulted in coat protein accumulation to a level 26% of that resulting from inoculation with transcripts with 25 adenylate residues (designated as wild type). Coat protein accumulation levels of 69 and 98% relative to wild type were observed after inoculation of protoplasts with transcripts bearing poly(A) tails 18 and 22 nucleotides long, respectively. The presence of a putative 3' pseudoknot structure including at least 13 adenylate residues of the 3'-terminal poly(A) tail was supported by enzymatic and chemical structural analysis. The functional relevance of this putative pseudoknot was tested by mutations that affected basepairing within the pseudoknot. These results support the existence of functional 3' pseudoknot that includes part of the 3' poly(A) tail.

* Corresponding author. Mailing address: Graduate Institute of Agricultural Biotechnology, National Chung Hsing University, Taichung 402, Taiwan. Phone: (886)-4-2840451. Fax: (886)-4-2860260. E-mail: chtsail{at}dragon.nchu.edu.tw.

Journal of Virology, April 1999, p. 2703-2709, Vol. 73, No. 4
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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