Human papillomavirus DNA replication compartments in a transient DNA replication system.
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.
Swindle
C S
CS
Departments of Biochemistry and Molecular Genetics, Schools of Medicine and Dentistry, University of Alabama at Birmingham, Birmingham, Alabama, USA.
Zou
N
N
Van Tine
B A
BA
Shaw
G M
GM
Engler
J A
JA
Chow
L T
LT
D002467:Cell Nucleus; D004261:DNA Replication; D004279:DNA, Viral; D004268:DNA-Binding Proteins / Q000032:analysis* / Q000235:genetics; D006801:Humans; D009363:Neoplasm Proteins / Q000032:analysis; D009687:Nuclear Proteins / Q000032:analysis; D027383:Papillomaviridae / Q000235:genetics*; D010957:Plasmids; D000071156:Promyelocytic Leukemia Protein; D018741:Replication Origin; D051716:Replication Protein A; D014157:Transcription Factors / Q000032:analysis; D014407:Tumor Cells, Cultured; D025521:Tumor Suppressor Proteins; D014764:Viral Proteins / Q000032:analysis* / Q000235:genetics; D014779:Virus Replication
D004279:DNA, Viral; D004268:DNA-Binding Proteins; C084304:E1 protein, Human papillomavirus type 11; D009363:Neoplasm Proteins; D009687:Nuclear Proteins; D000071156:Promyelocytic Leukemia Protein; C497167:RPA1 protein, human; D051716:Replication Protein A; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins; D014764:Viral Proteins; C070576:PML protein, human