Evidence that the first strand-transfer reaction of duck hepatitis B virus reverse transcription requires the polymerase and that strand transfer is not needed for the switch of the polymerase to the elongation mode of DNA synthesis.
Deletion of amino acids 79-88 in the duck hepatitis B virus reverse transcriptase had minimal effects on polymerase activities prior to the minus-strand DNA transfer reaction, yet it greatly diminished strand transfer and subsequent DNA synthesis. This mutation also reduced reverse transcription on exogenous RNA templates. The reaction on exogenous RNAs employed the phosphonoformic acid (PFA)-sensitive elongation mode of DNA synthesis rather than the PFA-resistant priming mode, despite the independence of DNA synthesis in this assay from the priming and minus-strand transfer reactions. These data provide experimental evidence that the polymerase is involved directly in the minus-strand transfer reaction and that the switch of the polymerase from the early PFA-resistant mode of DNA synthesis to the later PFA-sensitive elongation mode does not require the strand-transfer reaction.
Gong
Yunhao
Y
Department of Molecular Microbiology and Immunology, St Louis University School of Medicine, 1402 S. Grand Blvd, St Louis, MO 63104, USA1.
Yao
Ermei
E
Department of Molecular Microbiology and Immunology, St Louis University School of Medicine, 1402 S. Grand Blvd, St Louis, MO 63104, USA1.
Stevens
Melissa
M
Department of Molecular Microbiology and Immunology, St Louis University School of Medicine, 1402 S. Grand Blvd, St Louis, MO 63104, USA1.
Tavis
John E
JE
Department of Molecular Microbiology and Immunology, St Louis University School of Medicine, 1402 S. Grand Blvd, St Louis, MO 63104, USA1.
D004279:DNA, Viral / Q000096:biosynthesis*; D012321:DNA-Directed RNA Polymerases / Q000502:physiology*; D017245:Foscarnet / Q000494:pharmacology; D015611:Hepatitis B Virus, Duck / Q000235:genetics*; D009154:Mutation; D014158:Transcription, Genetic; D014779:Virus Replication
D004279:DNA, Viral; D017245:Foscarnet; D012321:DNA-Directed RNA Polymerases