title: Architecture of replication compartments formed during Epstein-Barr virus lytic replication.
abstract: Epstein-Barr virus (EBV) productive DNA replication occurs at discrete sites, called replication compartments, in nuclei. In this study we performed comprehensive analyses of the architecture of the replication compartments. The BZLF1 oriLyt binding proteins showed a fine, diffuse pattern of distribution throughout the nuclei at immediate-early stages of induction and then became associated with the replicating EBV genome in the replication compartments during lytic infection. The BMRF1 polymerase (Pol) processivity factor showed a homogenous, not dot-like, distribution in the replication compartments, which completely coincided with the newly synthesized viral DNA. Inhibition of viral DNA replication with phosphonoacetic acid, a viral DNA Pol inhibitor, eliminated the DNA-bound form of the BMRF1 protein, although the protein was sufficiently expressed in the cells. These observations together with the findings that almost all abundantly expressed BMRF1 proteins existed in the DNA-bound form suggest that the BMRF1 proteins not only act at viral replication forks as Pol processive factors but also widely distribute on newly replicated EBV genomic DNA. In contrast, the BALF5 Pol catalytic protein, the BALF2 single-stranded-DNA binding protein, and the BBLF2/3 protein, a component of the helicase-primase complex, were colocalized as distinct dots distributed within replication compartments, representing viral replication factories. Whereas cellular replication factories are constructed based on nonchromatin nuclear structures and nuclear matrix, viral replication factories were easily solubilized by DNase I treatment. Thus, compared with cellular DNA replication, EBV lytic DNA replication factories would be simpler so that construction of the replication domain would be more relaxed.
authors.affiliation: Division of Virology, Aichi Cancer Center Research Institute, 1-1, Kanokoden, Chikusa-ku, Nagoya 464-8681, Japan.
authors.forename: Tohru
authors.initials: T
authors.lastname: Daikoku
authors.forename: Ayumi
authors.initials: A
authors.lastname: Kudoh
authors.forename: Masatoshi
authors.initials: M
authors.lastname: Fujita
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Sugaya
authors.forename: Hiroki
authors.initials: H
authors.lastname: Isomura
authors.forename: Noriko
authors.initials: N
authors.lastname: Shirata
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Tsurumi
mesh_terms: D000956:Antigens, Viral / Q000378:metabolism; D002467:Cell Nucleus / Q000737:chemistry*; D004261:DNA Replication; D004279:DNA, Viral / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D003850:Deoxyribonuclease I / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005455:Fluorescent Antibody Technique; D004854:Herpesvirus 4, Human / Q000502:physiology*; D010746:Phosphonoacetic Acid / Q000494:pharmacology; D015534:Trans-Activators / Q000378:metabolism; D014764:Viral Proteins / Q000378:metabolism; D014779:Virus Replication
chemical_list: D000956:Antigens, Viral; C096364:BALF2 protein, Human herpesvirus 4; C063676:BZLF1 protein, Herpesvirus 4, Human; D004279:DNA, Viral; D004268:DNA-Binding Proteins; D004791:Enzyme Inhibitors; C083204:Epstein-Barr virus early antigen diffuse component; D015534:Trans-Activators; D014764:Viral Proteins; C083202:BALF5 protein, Epstein-barr virus; D004259:DNA-Directed DNA Polymerase; D003850:Deoxyribonuclease I; D010746:Phosphonoacetic Acid
references.citation: J Virol. 2004 May;78(9):4783-96
references.pmid: 15078960
references.citation: Virology. 2003 May 10;309(2):232-47
references.pmid: 12758171
references.citation: Cell. 1988 Nov 4;55(3):427-33
references.pmid: 2846181
references.citation: J Cell Sci. 1988 May;90 ( Pt 1):1-6
references.pmid: 3058728
references.citation: J Virol. 1991 Jan;65(1):483-8
references.pmid: 1845903
references.citation: Virology. 1991 Nov;185(1):309-15
references.pmid: 1656589
references.citation: J Virol. 1991 Dec;65(12):7073-7
references.pmid: 1658397
references.citation: J Virol. 1993 Mar;67(3):1681-7
references.pmid: 8382315
references.citation: Cell. 1993 Apr 23;73(2):361-73
references.pmid: 8097433
references.citation: J Virol. 1993 Aug;67(8):4651-8
references.pmid: 8392605
references.citation: J Virol. 1993 Dec;67(12):7648-53
references.pmid: 8230484
references.citation: J Virol. 1995 May;69(5):2998-3006
references.pmid: 7707526
references.citation: Int Rev Cytol. 1995;162A:1-65
references.pmid: 8575878
references.citation: Virology. 1996 Aug 15;222(2):352-64
references.pmid: 8806519
references.citation: Science. 1998 Apr 24;280(5363):547-53
references.pmid: 9554838
references.citation: J Biol Chem. 1998 Jul 3;273(27):17095-101
references.pmid: 9642275
references.citation: J Cell Biol. 1998 Dec 14;143(6):1415-25
references.pmid: 9852140
references.citation: Virology. 1998 Dec 5;252(1):162-78
references.pmid: 9875326
references.citation: J Biol Chem. 1999 Sep 3;274(36):25927-32
references.pmid: 10464337
references.citation: Virology. 1999 Sep 15;262(1):9-17
references.pmid: 10489336
references.citation: J Gen Virol. 1999 Nov;80 ( Pt 11):2879-87
references.pmid: 10580049
references.citation: J Virol. 2000 Mar;74(6):2550-7
references.pmid: 10684269
references.citation: J Cell Biol. 2000 Apr 17;149(2):271-80
references.pmid: 10769021
references.citation: EMBO J. 2000 Jun 15;19(12):3080-9
references.pmid: 10856251
references.citation: J Virol. 2000 Dec;74(24):11800-10
references.pmid: 11090180
references.citation: Curr Top Microbiol Immunol. 2001;258:65-87
references.pmid: 11443868
references.citation: J Biol Chem. 2002 Mar 22;277(12):10354-61
references.pmid: 11779870
references.citation: J Virol. 2003 Jan;77(2):851-61
references.pmid: 12502801
references.citation: EMBO J. 1986 Jun;5(6):1403-10
references.pmid: 3732249