A novel cis-acting element facilitates minus-strand DNA synthesis during reverse transcription of the hepatitis B virus genome

Myeong Kyun Shin, Jehan Lee, Wang-Shick Ryu

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

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 β5 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 β5 region diminished minus-strand DNA synthesis initiated at DR1*. Furthermore, the insertion of the β5 region into its own position in a mutant in which the sequences including the β5 region were replaced restored minus-strand DNA synthesis at DR1*. We speculate that the β5 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.

Original languageEnglish
Pages (from-to)6252-6262
Number of pages11
JournalJournal of Virology
Volume78
Issue number12
DOIs
Publication statusPublished - 2004 Jun 1

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reverse transcription
Hepatitis B virus
Reverse Transcription
Genome
Nucleotides
nucleotides
synthesis
genome
DNA
Hepadnaviridae
Sequence Inversion
circular DNA
Circular DNA
Base Pairing
RNA
mutants
stems
Proteins
proteins

All Science Journal Classification (ASJC) codes

  • Immunology

Cite this

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abstract = "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 β5 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 β5 region diminished minus-strand DNA synthesis initiated at DR1*. Furthermore, the insertion of the β5 region into its own position in a mutant in which the sequences including the β5 region were replaced restored minus-strand DNA synthesis at DR1*. We speculate that the β5 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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A novel cis-acting element facilitates minus-strand DNA synthesis during reverse transcription of the hepatitis B virus genome. / Shin, Myeong Kyun; Lee, Jehan; Ryu, Wang-Shick.

In: Journal of Virology, Vol. 78, No. 12, 01.06.2004, p. 6252-6262.

Research output: Contribution to journalArticle

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