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Journal of Virology, February 2004, p. 1129-1138, Vol. 78, No. 3
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.3.1129-1138.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Bunyamwera Bunyavirus RNA Synthesis Requires Cooperation of 3'- and 5'-Terminal Sequences

John N. Barr* and Gail W. Wertz

Department of Microbiology, University of Alabama School of Medicine, Birmingham, Alabama 35294

Received 27 August 2003/ Accepted 22 October 2003

Bunyamwera virus (BUNV) is the prototype of both the Orthobunyavirus genus and the Bunyaviridae family of segmented negative-sense RNA viruses. The tripartite BUNV genome consists of small (S), medium (M), and large (L) segments that are each transcribed to yield a single mRNA and are replicated to generate an antigenome that acts as a template for synthesis of further genomic strands. As for all negative-sense RNA viruses, the 3'- and 5'-terminal nontranslated regions (NTRs) of the BUNV S, M, and L segments exhibit nucleotide complementarity and, except for one conserved U-G pairing, this complementarity extends for 15, 18, and 19 nucleotides, respectively. We investigated whether the complementarity of 3' and 5' NTRs reflected a functional requirement for terminal cooperation to promote BUNV RNA synthesis or, alternatively, was a consequence of genomic and antigenomic NTRs having similar functions requiring sequence conservation. We show that cooperation between 3'- and 5'-NTR sequences is required for BUNV RNA synthesis, and our results suggest that this cooperation is due to nucleotide complementarity allowing 3' and 5' NTRs to associate through base-pairing interactions. To examine the importance of complementarity in promoting BUNV RNA synthesis, we utilized a competitive replication assay able to examine the replication ability of all possible combinations of interacting nucleotides within a defined region of BUNV 3' and 5' NTRs. We show here that maximal RNA replication was signaled when sequences exhibiting perfect complementarity within 3' and 5' NTRs were selected.

* Corresponding author. Mailing address: Department of Microbiology, University of Alabama School of Medicine, BBRB Rm. 360/17, 845 19th St. South, Birmingham, AL 35294-2170. Phone: (205) 934-0453. Fax: (205) 934-1636. E-mail: jbarr{at}uab.edu.

Journal of Virology, February 2004, p. 1129-1138, Vol. 78, No. 3
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.3.1129-1138.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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