Architecture of replication compartments formed during Epstein-Barr virus lytic replication.
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.
Daikoku
Tohru
T
Division of Virology, Aichi Cancer Center Research Institute, 1-1, Kanokoden, Chikusa-ku, Nagoya 464-8681, Japan.
Kudoh
Ayumi
A
Fujita
Masatoshi
M
Sugaya
Yutaka
Y
Isomura
Hiroki
H
Shirata
Noriko
N
Tsurumi
Tatsuya
T
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
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
J Virol. 2004 May;78(9):4783-96
15078960
Virology. 2003 May 10;309(2):232-47
12758171
Cell. 1988 Nov 4;55(3):427-33
2846181
J Cell Sci. 1988 May;90 ( Pt 1):1-6
3058728
J Virol. 1991 Jan;65(1):483-8
1845903
Virology. 1991 Nov;185(1):309-15
1656589
J Virol. 1991 Dec;65(12):7073-7
1658397
J Virol. 1993 Mar;67(3):1681-7
8382315
Cell. 1993 Apr 23;73(2):361-73
8097433
J Virol. 1993 Aug;67(8):4651-8
8392605
J Virol. 1993 Dec;67(12):7648-53
8230484
J Virol. 1995 May;69(5):2998-3006
7707526
Int Rev Cytol. 1995;162A:1-65
8575878
Virology. 1996 Aug 15;222(2):352-64
8806519
Science. 1998 Apr 24;280(5363):547-53
9554838
J Biol Chem. 1998 Jul 3;273(27):17095-101
9642275
J Cell Biol. 1998 Dec 14;143(6):1415-25
9852140
Virology. 1998 Dec 5;252(1):162-78
9875326
J Biol Chem. 1999 Sep 3;274(36):25927-32
10464337
Virology. 1999 Sep 15;262(1):9-17
10489336
J Gen Virol. 1999 Nov;80 ( Pt 11):2879-87
10580049
J Virol. 2000 Mar;74(6):2550-7
10684269
J Cell Biol. 2000 Apr 17;149(2):271-80
10769021
EMBO J. 2000 Jun 15;19(12):3080-9
10856251
J Virol. 2000 Dec;74(24):11800-10
11090180
Curr Top Microbiol Immunol. 2001;258:65-87
11443868
J Biol Chem. 2002 Mar 22;277(12):10354-61
11779870
J Virol. 2003 Jan;77(2):851-61
12502801
EMBO J. 1986 Jun;5(6):1403-10
3732249