Sequence identity of the direct repeats, DR1 and DR2, contributes to the discrimination between primer translocation and in situ priming during replication of the duck hepatitis B virus
| Autor: | Jeffrey W. Habig, Daniel D. Loeb |
|---|---|
| Rok vydání: | 2006 |
| Předmět: |
DNA Replication
DNA polymerase Molecular Sequence Data chemical and pharmacologic phenomena Virus Replication Article Cell Line Hepatitis B Virus Duck chemistry.chemical_compound Structural Biology Animals RNase H Molecular Biology Repetitive Sequences Nucleic Acid biology DNA replication Molecular biology Reverse transcriptase Ducks chemistry biology.protein Nucleic Acid Conformation Primase Primer (molecular biology) Chickens DNA In vitro recombination |
| Zdroj: | Journal of molecular biology. 364(1) |
| ISSN: | 0022-2836 |
| Popis: | There are two mutually exclusive pathways for plus-strand DNA synthesis in hepadnavirus reverse transcription. The predominant pathway gives rise to relaxed circular DNA, while the other pathway yields duplex linear DNA. At the completion of minus-strand DNA synthesis, the final RNase H cleavage generates the plus-strand primer at direct repeat 1 (DR1). A small fraction of viruses make duplex linear DNA after initiating plus-strand DNA synthesis from this site, a process called in situ priming. To make relaxed circular DNA, a template switch is necessary for the RNA primer generated at DR1 to initiate plus-strand DNA synthesis from the direct repeat 2 (DR2) located near the opposite end of the minus-strand DNA, a process called primer translocation. We are interested in understanding the mechanism that discriminates between these two processes. Previously, we showed that a small DNA hairpin forms at DR1 in the avihepadnaviruses and acts as an inhibitor of in situ priming. Here, using genetic approaches, we show that sequence identity between DR1 and DR2 is necessary, but not sufficient for primer translocation in the duck hepatitis B virus. The discrimination between in situ priming and primer translocation depends upon suppression of in situ priming, a process that is dependent upon both sequence identity between DR1 and DR2, and the presence of the hairpin at DR1. Finally, our analysis indicates the entire RNA primer can contribute to primer translocation and is translocated to DR2 before initiation of plus-strand DNA synthesis from that site. |
| Databáze: | OpenAIRE |
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