Formation of herpes simplex virus type 1 replication compartments by transfection: requirements and localization to nuclear domain 10.
During infection, the seven essential herpes simplex virus type 1 (HSV-1) replication proteins are found in globular nuclear structures called replication compartments. Replication compartments form adjacent to ND10, nuclear matrix-bound domains which are present in most cell types but whose function is unknown (G. G. Maul, I. M. Ishov, and R. D. Everett, Virology 217:67-75, 1996). We now demonstrate that replication compartments can be formed by cotransfecting Vero cells with constructs expressing the seven essential viral replication proteins and a plasmid containing an HSV-1 origin of DNA replication. Like replication compartments in infected cells, replication compartments formed by cotransfection contain all of the essential viral replication proteins, are sites of DNA synthesis, and are found adjacent to ND10. However, neither the viral origin-binding protein nor a plasmid containing an HSV-1 origin of DNA replication is individually required for the formation of transfection replication compartments, although the presence of each increases the efficiency of replication compartment formation. Further, we provide evidence that UL29 independently localizes adjacent to ND10 and so may play a role in directing replication compartments to these preexisting nuclear structures.
Lukonis
C J
CJ
Department of Microbiology, University of Connecticut Health Center, Farmington 06030, USA.
Weller
S K
SK
D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D002522:Chlorocebus aethiops; D004265:DNA Helicases / Q000235:genetics / Q000378:metabolism*; D019915:DNA Primase; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D004259:DNA-Directed DNA Polymerase / Q000235:genetics / Q000378:metabolism*; D005090:Exodeoxyribonucleases; D018259:Herpesvirus 1, Human / Q000378:metabolism* / Q000502:physiology; D006801:Humans; D009687:Nuclear Proteins / Q000378:metabolism; D010957:Plasmids; D018741:Replication Origin; D014162:Transfection; D014709:Vero Cells; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication
D004268:DNA-Binding Proteins; C045462:ICP8 protein, Simplexvirus; D009687:Nuclear Proteins; D014764:Viral Proteins; C064565:UL9 protein, Human herpesvirus 1; D019915:DNA Primase; D004259:DNA-Directed DNA Polymerase; D005090:Exodeoxyribonucleases; C062021:helicase-primase, Human herpesvirus 1; C043417:DNA polymerase, Simplexvirus; D004265:DNA Helicases