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Journal of Virology, January 2003, p. 245-257, Vol. 77, No. 1
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.1.245-257.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

An Internally Located RNA Hairpin Enhances Replication of Tomato Bushy Stunt Virus RNAs

Debashish Ray and K. Andrew White*

Department of Biology, York University, Toronto, Ontario, Canada M3J 1P3

Received 28 June 2002/ Accepted 24 September 2002

Defective interfering (DI) RNAs of Tomato bushy stunt virus (TBSV), a plus-sense RNA virus, comprise four conserved noncontiguous regions (I through IV) derived from the viral genome. Region III, a 70-nucleotide-long sequence corresponding to a genomic segment located 378 nucleotides upstream of the 3' terminus of the genome, has been found to enhance DI RNA accumulation by approximately 10-fold in an orientation-independent manner (D. Ray and K. A. White, Virology 256:162-171, 1999). In this study, a more detailed structure-function analysis of region III was conducted. RNA secondary-structure analyses indicated that region III contains stem-loop structures in both plus and minus strands. Through deletion analyses of a DI RNA, a primary determinant of region III activity was mapped to the 5'-proximal 35-nucleotide segment. Compensatory-type mutational analyses showed that a stem-loop structure in the minus strand of this subregion was required for enhanced DI RNA replication. The same stem-loop structure was also found to function in a position-independent manner in a DI RNA (albeit at reduced levels) and to be important for efficient accumulation within the context of the TBSV genome. Taken together, these observations suggest that the 5'-proximal segment of region III is a modular RNA replication element that functions primarily through the formation of an RNA hairpin structure in the minus strand.

* Corresponding author. Mailing address: Department of Biology, York University, 4700 Keele St., Toronto, Ontario, Canada M3J 1P3. Phone: (416) 736-2100 (ext. 40890 or 70352). Fax: (416) 736-5698. E-mail: kawhite{at}yorku.ca.

Journal of Virology, January 2003, p. 245-257, Vol. 77, No. 1
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.1.245-257.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Panavas, T., Nagy, P. D. (2002). The RNA Replication Enhancer Element of Tombusviruses Contains Two Interchangeable Hairpins That Are Functional during Plus-Strand Synthesis. J. Virol. 77: 258-269 [Abstract] [Full Text]  


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