Kinetics of early molecular events in duck hepatitis B virus replication in primary duck hepatocytes.
This paper describes the use of one-step growth conditions to study the kinetics of duck hepatitis B virus (DHBV) replication in primary duck hepatocytes. Synchronized infection was achieved using partially purified DHBV virions at an m.o.i. of 640 DHBV DNA-containing virions per cell, and these conditions were shown to produce a single cycle of infection. In this model, input purified DHBV DNA was rapidly internalized by cells at > or = 0.5 h, and localized to the nucleus by 4 h, but both covalently closed circular (CCC) DNA and single-stranded DNA were not detected until 48 h postinoculation (p.i.), suggesting that there was a > or = 40 h delay between DHBV localization to the nucleus and formation of CCC DNA. In contrast, CCC DNA can be first detected in hepatocytes at 6 h p.i. in in vivo infection of ducks with the same DHBV strain. In an analysis of the nuclear transport of the DHBV genome, release of nuclear viral DNA from a particulate form to a soluble nucleoplasmic form was only 50% complete by 48 h p.i. However, this process occurred simultaneously with genome uncoating since all soluble nucleoplasmic DHBV DNA was free of nucleocapsid material; this suggests that nucleocapsid disassembly and genome uncoating may occur at the nuclear membrane and not within the nucleus. Quantitative analysis demonstrated inefficiency in a number of steps including virus uptake and internalization, translocation of nucleocapsid across the nuclear membrane and antigen expression from intranuclear viral DNA.
Qiao
M
M
Infectious Diseases Laboratories, Institute of Medical and Veterinary Science, PO Box 14, Rundle Mall, Adelaide, SA 5000, Australia1.
Scougall
C A
CA
Department of Microbiology and Immunology, University of Adelaide, Adelaide, SA 5005, Australia2.
Duszynski
A
A
Department of Microbiology and Immunology, University of Adelaide, Adelaide, SA 5005, Australia2.
Burrell
C J
CJ
Department of Microbiology and Immunology, University of Adelaide, Adelaide, SA 5005, Australia2.|Infectious Diseases Laboratories, Institute of Medical and Veterinary Science, PO Box 14, Rundle Mall, Adelaide, SA 5000, Australia1.
D000818:Animals; D001692:Biological Transport; D002467:Cell Nucleus / Q000821:virology; D002478:Cells, Cultured; D004261:DNA Replication; D004279:DNA, Viral / Q000378:metabolism; D004372:Ducks; D015611:Hepatitis B Virus, Duck / Q000235:genetics / Q000254:growth & development / Q000502:physiology*; D007700:Kinetics; D008099:Liver / Q000166:cytology / Q000821:virology*; D014779:Virus Replication / Q000235:genetics
D004279:DNA, Viral