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Journal of Virology, February 1999, p. 1001-1009, Vol. 73, No. 2
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Human Papillomavirus DNA Replication Compartments in a Transient DNA Replication System

C. Scott Swindle,1 Nianxiang Zou,1 Brian A. Van Tine,2,3 George M. Shaw,2,4 Jeffrey A. Engler,1,* and Louise T. Chow1,*

Departments of Biochemistry and Molecular Genetics,1 Medicine,2 and Pathology3 and The Howard Hughes Medical Institute,4 Schools of Medicine and Dentistry, University of Alabama at Birmingham, Birmingham, Alabama

Received 26 May 1998/Accepted 28 October 1998

Many DNA viruses replicate their genomes at nuclear foci in infected cells. Using indirect immunofluorescence in combination with fluorescence in situ hybridization, we colocalized the human papillomavirus (HPV) replicating proteins E1 and E2 and the replicating origin-containing plasmid to nuclear foci in transiently transfected cells. The host replication protein A (RP-A) was also colocalized to these foci. These nuclear structures were identified as active sites of viral DNA synthesis by bromodeoxyuridine (BrdU) pulse-labeling. Unexpectedly, the great majority of RP-A and BrdU incorporation was found in these HPV replication domains. Furthermore, E1, E2, and RP-A were also colocalized to nuclear foci in the absence of an origin-containing plasmid. These observations suggest a spatial reorganization of the host DNA replication machinery upon HPV DNA replication or E1 and E2 expression. Alternatively, viral DNA replication might be targeted to host nuclear domains that are active during the late S phase, when such domains are limited in number. In a fraction of cells expressing E1 and E2, the promyelocytic leukemia protein, a component of nuclear domain 10 (ND10), was either partially or completely colocalized with E1 and E2. Since ND10 structures were recently hypothesized to be sites of bovine papillomavirus virion assembly, our observation suggests that HPV DNA amplification might be partially coupled to virion assembly.

* Corresponding author. Mailing address for Jeffrey A. Engler: University of Alabama at Birmingham, Department of Biochemistry and Molecular Genetics, 1918 University Blvd., Birmingham, AL 35294-0005. Phone: (205) 934-4734. Fax: (205) 934-0758. E-mail: jengler{at}uab.edu. Mailing address for Louise T. Chow: University of Alabama at Birmingham, Department of Biochemistry and Molecular Genetics, 1918 University Blvd., Birmingham, AL 35294-0005. Phone: (205) 975-8300. Fax: (205) 975-6075. E-mail: ltchow{at}uab.edu.

Journal of Virology, February 1999, p. 1001-1009, Vol. 73, No. 2
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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