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Cell. 1986 Nov 7;47(3):451-60.

Formation of the pool of covalently closed circular viral DNA in hepadnavirus-infected cells.


Covalently closed circular (CCC) double-stranded DNA believed to be the transcriptional template for duck hepatitis B virus (DHBV) is amplified in aging primary cultures of hepatocytes from congenitally infected ducklings. Analysis of 5-bromodeoxyuridine-labeled heavy/light CCC DNA shows that the relaxed circular DNA synthesized in the cytoplasm by reverse transcription is the predominant precursor to the amplified pool of nuclear viral CCC DNA. In vitro infection of uninfected hepatocyte cultures with DHBV demonstrates that a similar 50-fold amplification of CCC DNA occurs during an early stage in the infection before virus production. This amplification allows the establishment of a pool of transcriptional templates in the cell without the need for semiconservative replication or multiple rounds of infection. This process may account for the ability of hepadnavirus-infected cells persistently to produce virus particles in the absence of stable integration of viral DNA.

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