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A Novel cis-Acting Element Facilitates Minus-Strand DNA Synthesis during Reverse Transcription of the Hepatitis B Virus Genome

Department of Biochemistry, Yonsei University, Seoul 120-749, Korea

Received 16 September 2003/ Accepted 29 February 2004

Hepadnaviruses replicate through reverse transcription of an RNA pregenome, resulting in a relaxed circular DNA genome. The first 3 or 4 nucleotides (nt) of minus-strand DNA are synthesized by the use of a bulge in a stem-loop structure near the 5' end of the pregenome as a template. This primer is then transferred to a complementary UUCA motif, termed an acceptor, within DR1* near the 3' end of the viral pregenome via 4-nt homology, and it resumes minus-strand DNA synthesis: this process is termed minus-strand transfer or primer translocation. Aside from the sequence identity of the donor and acceptor, little is known about the sequence elements contributing to minus-strand transfer. Here we report a novel cis-acting element, termed the ß5 region (28 nt in length), located 20 nt upstream of DR1*, that facilitates minus-strand DNA synthesis. The deletion or inversion of the sequence including the ß5 region diminished minus-strand DNA synthesis initiated at DR1*. Furthermore, the insertion of the ß5 region into its own position in a mutant in which the sequences including the ß5 region were replaced restored minus-strand DNA synthesis at DR1*. We speculate that the ß5 region facilitates minus-strand transfer, possibly by bringing the acceptor site in proximity to the donor site via base pairing or by interacting with protein factors involved in this process.

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• Abraham, T. M., Loeb, D. D. (2006). 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. J. Virol. 80: 4380-4387 [Abstract] [Full Text]