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Human papillomavirus DNA replication compartments in a transient DNA replication system
C S Swindle1, N Zou, B A Van Tine
1Departments of Biochemistry and Molecular Genetics, Schools of Medicine and Dentistry, University of Alabama at Birmingham, Birmingham, Alabama, USA.
Journal of Virology
|January 9, 1999
Summary
Human papillomavirus (HPV) replication proteins E1 and E2 recruit host replication protein A (RP-A) to nuclear foci, forming active viral DNA synthesis sites. This suggests HPV reorganizes host DNA replication machinery or targets late S-phase domains.
Area of Science:
- Molecular Biology
- Virology
- Cell Biology
Background:
- DNA viruses often replicate genomes within nuclear foci.
- Human papillomavirus (HPV) replication involves specific viral proteins and host factors.
Purpose of the Study:
- To investigate the spatial organization of HPV DNA replication machinery within infected cells.
- To determine the role of host replication protein A (RP-A) in HPV DNA synthesis.
- To explore the potential link between HPV replication and nuclear domain structures.
Main Methods:
- Indirect immunofluorescence and fluorescence in situ hybridization (FISH) to colocalize viral proteins (E1, E2), origin-containing plasmids, and host protein RP-A.
- Bromodeoxyuridine (BrdU) pulse-labeling to identify active DNA synthesis sites.
- Analysis of promyelocytic leukemia protein and nuclear domain 10 (ND10) colocalization.
Main Results:
- HPV replication proteins E1 and E2, along with origin-containing plasmids, localize to nuclear foci.
- Host replication protein A (RP-A) colocalizes with E1 and E2 at these foci, which are active sites of viral DNA synthesis.
- A significant portion of RP-A and BrdU incorporation occurs within HPV replication domains, indicating recruitment of host machinery.
- E1, E2, and RP-A also form foci independently of the origin-containing plasmid, suggesting protein-driven reorganization.
- Colocalization with promyelocytic leukemia protein suggests a potential link between HPV DNA amplification and nuclear domain 10 (ND10) structures.
Conclusions:
- HPV DNA replication induces a spatial reorganization of the host DNA replication machinery.
- Viral replication may target host nuclear domains active during late S-phase.
- HPV DNA amplification might be partially coupled to virion assembly at ND10 sites.