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Journal of Virology, May 2006, p. 4380-4387, Vol. 80, No. 9
0022-538X/06/$08.00+0     doi:10.1128/JVI.80.9.4380-4387.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Base Pairing between the 5' Half of {varepsilon} and a cis-Acting Sequence, {Phi}, Makes a Contribution to the Synthesis of Minus-Strand DNA for Human Hepatitis B Virus

Teresa M. Abraham and Daniel D. Loeb*

McArdle Laboratory for Cancer Research, University of Wisconsin Medical School, Madison, Wisconsin 53706

Received 29 November 2005/ Accepted 9 February 2006

Synthesis of minus-strand DNA of human hepatitis B virus (HBV) can be divided into three phases: initiation of DNA synthesis, the template switch, and elongation of minus-strand DNA. Although much is known about minus-strand DNA synthesis, the mechanism(s) by which this occurs has not been completely elucidated. Through a deletion analysis, we have identified a cis-acting element involved in minus-strand DNA synthesis that lies within a 27-nucleotide region between DR2 and the 3' copy of DR1. A subset of this region (termed {Phi}) has been hypothesized to base pair with the 5' half of {varepsilon} (H. Tang and A. McLachlan, Virology, 303:199-210, 2002). To test the proposed model, we used a genetic approach in which multiple sets of variants that disrupted and then restored putative base pairing between the 5' half of {varepsilon} and {Phi} were analyzed. Primer extension analysis, using two primers simultaneously, was performed to measure encapsidated pregenomic RNA (pgRNA) and minus-strand DNA synthesized in cell culture. The efficiency of minus-strand DNA synthesis was defined as the amount of minus-strand DNA synthesized per encapsidation event. Our results indicate that base pairing between {Phi} and the 5' half of {varepsilon} contributes to efficient minus-strand DNA synthesis. Additional results are consistent with the idea that the primary sequence of {Phi} and/or {varepsilon} also contributes to function. How base pairing between {Phi} and {varepsilon} contributes to minus-strand DNA synthesis is not known, but a simple speculation is that {Phi} base pairs with the 5' half of {varepsilon} to juxtapose the donor and acceptor sites to facilitate the first-strand template switch.

* Corresponding author. Mailing address: McArdle Laboratory for Cancer Research, University of Wisconsin Medical School, 1400 University Ave., Madison, WI 53706. Phone: (608) 262-1260. Fax: (608) 262-2824. E-mail: loeb{at}oncology.wisc.edu.

Journal of Virology, May 2006, p. 4380-4387, Vol. 80, No. 9
0022-538X/06/$08.00+0     doi:10.1128/JVI.80.9.4380-4387.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Wang, H., Kim, S., Ryu, W.-S. (2009). DDX3 DEAD-Box RNA Helicase Inhibits Hepatitis B Virus Reverse Transcription by Incorporation into Nucleocapsids. J. Virol. 83: 5815-5824 [Abstract] [Full Text]  
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  • Abraham, T. M., Loeb, D. D. (2007). The Topology of Hepatitis B Virus Pregenomic RNA Promotes Its Replication. J. Virol. 81: 11577-11584 [Abstract] [Full Text]  
  • Lewellyn, E. B., Loeb, D. D. (2007). Base Pairing between cis-Acting Sequences Contributes to Template Switching during Plus-Strand DNA Synthesis in Human Hepatitis B Virus. J. Virol. 81: 6207-6215 [Abstract] [Full Text]  


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