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PMID: 11152521 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Origin-independent assembly of Kaposi's sarcoma-associated herpesvirus DNA replication compartments in transient cotransfection assays and association with the ORF-K8 protein and cellular PML.

Journal of virology ·Vol. 75 ·No. 3 ·2001-02-00 ·Pages 1487-506

Wu FY, Ahn JH, Alcendor DJ, Jang WJ, Xiao J, Hayward SD, Hayward GS

Abstract

Six predicted Kaposi's sarcoma virus herpesvirus (KSHV) proteins have homology with other well-characterized herpesvirus core DNA replication proteins and are expected to be essential for viral DNA synthesis. Intact Flag-tagged protein products from all six were produced from genomic expression vectors, although the ORF40/41 transcript encoding a primase-helicase component proved to be spliced with a 127-bp intron. The intracellular localization of these six KSHV replication proteins and the mechanism of their nuclear translocation were investigated. SSB (single-stranded DNA binding protein, ORF6) and PPF (polymerase processivity factor, ORF59) were found to be intrinsic nuclear proteins, whereas POL (polymerase, ORF9), which localized in the cytoplasm on its own, was translocated to the nucleus when cotransfected with PPF. PAF (primase-associated factor, ORF40/41), a component of the primase-helicase tripartite subcomplex together with PRI (primase, ORF56) and HEL (helicase, ORF44), required the presence of all five other replication proteins for efficient nuclear translocation. Surprisingly, even in the absence of a lytic cycle replication origin (ori-Lyt) and any known initiator or origin binding protein, the protein products of all six KSHV core replication genes cooperated in a transient cotransfection assay to form large globular shaped pseudo-replication compartments (pseudo-RC), which excluded cellular DNA. These pseudo-RC structures were confirmed to include POL, SSB, PRI, and PAF but did not contain any newly synthesized DNA. Similar to the human cytomegalovirus system, the peripheries of these KSHV pre-RC were also found to be surrounded by punctate PML oncogenic domains (PODs). Furthermore, by transient cotransfection, the six KSHV core replication machinery proteins successfully replicated a plasmid containing EBV ori-Lyt in the presence of the Epstein-Barr virus-encoded DNA binding initiator protein, ZTA. The KSHV-encoded K8 (ORF-K8) protein, which is a distant evolutionary homologue to ZTA, was incorporated into pseudo-RC structures formed by transient cotransfection with the six core KSHV replication genes. However, unlike ZTA, K8 displayed a punctate nuclear pattern both in transfected cells and at early stages of lytic infection and colocalized with the cellular PML proteins in PODs. Finally, K8 was also found to accumulate in functional viral RC, detected by incorporation of pulse-labeled bromodeoxyuridine into newly synthesized DNA in both tetradecanoyl phorbol acetate-induced JSC-1 primary effusion lymphoblasts and in KSHV lytically infected endothelial cells.

MeSH Terms
Amino Acid Sequence Animals Antifungal Agents Cell Nucleus/metabolism Chlorocebus aethiops Cloning, Molecular DNA Replication DNA-Binding Proteins/physiology Fungal Proteins/genetics,physiology Herpesvirus 8, Human/physiology Molecular Sequence Data Neoplasm Proteins/physiology Nuclear Proteins Open Reading Frames Penicillium Trans-Activators/physiology Transcription Factors/physiology Transfection Tumor Suppressor Proteins Vero Cells Viral Core Proteins/physiology Viral Proteins Virus Assembly Virus Replication
Chemicals
Antifungal Agents BZLF1 protein, Herpesvirus 4, Human DNA-Binding Proteins Fungal Proteins Neoplasm Proteins Nuclear Proteins PAF protein, Penicillium chrysogenum Trans-Activators Transcription Factors Tumor Suppressor Proteins Viral Core Proteins Viral Proteins
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Wu F Y
Molecular Virology Laboratories, Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, Maryland 21231-1000, USA.
Ahn J H
Alcendor D J
Jang W J
Xiao J
Hayward S D
Hayward G S
References (78)
78 references, click to expand
  1. The human cytomegalovirus IE2 and UL112-113 proteins accumulate in viral DNA replication compartments that initiate from the periphery of promyelocytic leukemia protein-associated nuclear bodies (PODs or ND10).
    J Virol. 1999 Dec;73(12):10458-71 PMID: 10559364
  2. Human herpesvirus-8-encoded LNA-1 accumulates in heterochromatin- associated nuclear bodies.
    J Gen Virol. 1999 Nov;80 ( Pt 11):2889-900 PMID: 10580050
  3. Review: properties and assembly mechanisms of ND10, PML bodies, or PODs.
    J Struct Biol. 2000 Apr;129(2-3):278-87 PMID: 10806078
  4. Kaposi's sarcoma-associated herpesvirus open reading frame 50/Rta protein activates the entire viral lytic cycle in the HH-B2 primary effusion lymphoma cell line.
    J Virol. 2000 Jul;74(13):6207-12 PMID: 10846108
  5. A new primary effusion lymphoma-derived cell line yields a highly infectious Kaposi's sarcoma herpesvirus-containing supernatant.
    J Virol. 2000 Nov;74(21):10187-93 PMID: 11024147
  6. A cis-acting element from the Epstein-Barr viral genome that permits stable replication of recombinant plasmids in latently infected cells.
    Proc Natl Acad Sci U S A. 1984 Jun;81(12):3806-10 PMID: 6328526
  7. Activation of expression of latent Epstein-Barr herpesvirus after gene transfer with a small cloned subfragment of heterogeneous viral DNA.
    Proc Natl Acad Sci U S A. 1985 Jun;82(12):4085-9 PMID: 2987963
  8. Both Epstein-Barr virus (EBV)-encoded trans-acting factors, EB1 and EB2, are required to activate transcription from an EBV early promoter.
    EMBO J. 1986 Dec 1;5(12):3243-9 PMID: 3028777
  9. Disruption of PML subnuclear domains by the acidic IE1 protein of human cytomegalovirus is mediated through interaction with PML and may modulate a RING finger-dependent cryptic transactivator function of PML.
    Mol Cell Biol. 1998 Aug;18(8):4899-913 PMID: 9671498
  10. A viral gene that activates lytic cycle expression of Kaposi's sarcoma-associated herpesvirus.
    Proc Natl Acad Sci U S A. 1998 Sep 1;95(18):10866-71 PMID: 9724796
  11. The Epstein-Barr virus Rta protein activates lytic cycle genes and can disrupt latency in B lymphocytes.
    J Virol. 1998 Oct;72(10):7978-84 PMID: 9733836
  12. Herpes simplex virus type 1 ICP27 is an essential regulatory protein.
    J Virol. 1985 Sep;55(3):796-805 PMID: 2991596
  13. Sequence-specific DNA binding of the Epstein-Barr virus nuclear antigen (EBNA-1) to clustered sites in the plasmid maintenance region.
    Cell. 1985 Oct;42(3):859-68 PMID: 2996781
  14. A method for identifying the viral genes required for herpesvirus DNA replication.
    Proc Natl Acad Sci U S A. 1986 Dec;83(23):9094-8 PMID: 3024166
  15. Localization of the herpes simplex virus type 1 65-kilodalton DNA-binding protein and DNA polymerase in the presence and absence of viral DNA synthesis.
    J Virol. 1990 Dec;64(12):5738-49 PMID: 2173766
  16. Correct intranuclear localization of herpes simplex virus DNA polymerase requires the viral ICP8 DNA-binding protein.
    J Virol. 1991 Mar;65(3):1082-9 PMID: 1847437
  17. ZEBRA and a Fos-GCN4 chimeric protein differ in their DNA-binding specificities for sites in the Epstein-Barr virus BZLF1 promoter.
    J Virol. 1991 Aug;65(8):4033-41 PMID: 1649314
  18. Cooperation of EBV DNA polymerase and EA-D(BMRF1) in vitro and colocalization in nuclei of infected cells.
    Virology. 1991 Sep;184(1):330-40 PMID: 1651595
  19. Transcriptional synergy by the Epstein-Barr virus transactivator ZEBRA.
    J Virol. 1992 Aug;66(8):4803-13 PMID: 1321270
  20. trans-acting requirements for replication of Epstein-Barr virus ori-Lyt.
    J Virol. 1992 Aug;66(8):5030-9 PMID: 1321285
  21. The Epstein-Barr virus R transactivator (Rta) contains a complex, potent activation domain with properties different from those of VP16.
    J Virol. 1992 Sep;66(9):5500-8 PMID: 1323708
  22. cis-acting elements in the lytic origin of DNA replication of Epstein-Barr virus.
    J Virol. 1993 Jul;67(7):4237-45 PMID: 8389925
  23. A transcription factor with homology to the AP-1 family links RNA transcription and DNA replication in the lytic cycle of Epstein-Barr virus.
    EMBO J. 1993 Oct;12(10):3921-9 PMID: 8404860
  24. Eleven loci encoding trans-acting factors are required for transient complementation of human cytomegalovirus oriLyt-dependent DNA replication.
    J Virol. 1993 Dec;67(12):6979-88 PMID: 8230421
  25. Further characterization of the interaction between the Epstein-Barr virus DNA polymerase catalytic subunit and its accessory subunit with regard to the 3'-to-5' exonucleolytic activity and stability of initiation complex at primer terminus.
    J Virol. 1994 May;68(5):3354-63 PMID: 8151794
  26. Interaction of herpes simplex virus type 1 DNA polymerase and the UL42 accessory protein with a model primer template.
    J Virol. 1994 Aug;68(8):4937-45 PMID: 8035492
  27. A mechanism for TAFs in transcriptional activation: activation domain enhancement of TFIID-TFIIA--promoter DNA complex formation.
    Genes Dev. 1994 May 1;8(9):995-1006 PMID: 7926793
  28. Evaluation of colocalization interactions between the IE110, IE175, and IE63 transactivator proteins of herpes simplex virus within subcellular punctate structures.
    J Virol. 1995 Jan;69(1):476-91 PMID: 7983744
  29. Identification of herpesvirus-like DNA sequences in AIDS-associated Kaposi's sarcoma.
    Science. 1994 Dec 16;266(5192):1865-9 PMID: 7997879
  30. Cytosolic factors in nuclear transport: what's importin?
    Cell. 1994 Dec 16;79(6):931-4 PMID: 8001141
  31. Bipartite DNA-binding region of the Epstein-Barr virus BMRF1 product essential for DNA polymerase accessory function.
    J Virol. 1995 Mar;69(3):1669-77 PMID: 7853503
  32. Kaposi's sarcoma-associated herpesvirus-like DNA sequences in AIDS-related body-cavity-based lymphomas.
    N Engl J Med. 1995 May 4;332(18):1186-91 PMID: 7700311
  33. Replication of Epstein-Barr virus oriLyt: lack of a dedicated virally encoded origin-binding protein and dependence on Zta in cotransfection assays.
    J Virol. 1995 May;69(5):2998-3006 PMID: 7707526
  34. Efficient persistence of extrachromosomal KSHV DNA mediated by latency-associated nuclear antigen.
    Science. 1999 Apr 23;284(5414):641-4 PMID: 10213686
  35. Identification of the immediate-early transcripts of Kaposi's sarcoma-associated herpesvirus.
    J Virol. 1999 Jul;73(7):5556-67 PMID: 10364304
  36. Lytic growth of Kaposi's sarcoma-associated herpesvirus (human herpesvirus 8) in culture.
    Nat Med. 1996 Mar;2(3):342-6 PMID: 8612236
  37. Genetic relations between varicella-zoster virus and Epstein-Barr virus.
    J Gen Virol. 1987 Apr;68 ( Pt 4):1067-79 PMID: 3033126
  38. Association of Epstein-Barr virus early antigen diffuse component and virus-specified DNA polymerase activity.
    J Virol. 1987 Sep;61(9):2947-9 PMID: 3039183
  39. Identification of herpes simplex virus type 1 genes required for origin-dependent DNA synthesis.
    J Virol. 1988 Feb;62(2):435-43 PMID: 2826806
  40. Structures of herpes simplex virus type 1 genes required for replication of virus DNA.
    J Virol. 1988 Feb;62(2):444-53 PMID: 2826807
  41. Herpes simplex virus DNA replication: the UL9 gene encodes an origin-binding protein.
    Proc Natl Acad Sci U S A. 1988 Aug;85(15):5414-8 PMID: 2840659
  42. Identification and characterization of oriLyt, a lytic origin of DNA replication of Epstein-Barr virus.
    Cell. 1988 Nov 4;55(3):427-33 PMID: 2846181
  43. Herpes simplex virus type 1 gene products required for DNA replication: identification and overexpression.
    J Virol. 1989 Jan;63(1):196-204 PMID: 2535726
  44. Epstein-Barr virus BZLF1 trans-activator specifically binds to a consensus AP-1 site and is related to c-fos.
    EMBO J. 1989 Jan;8(1):127-32 PMID: 2540954
  45. The Epstein-Barr virus BMLF1 promoter contains an enhancer element that is responsive to the BZLF1 and BRLF1 transactivators.
    J Virol. 1989 Sep;63(9):3878-83 PMID: 2548003
  46. The zta transactivator involved in induction of lytic cycle gene expression in Epstein-Barr virus-infected lymphocytes binds to both AP-1 and ZRE sites in target promoter and enhancer regions.
    J Virol. 1990 Mar;64(3):1143-55 PMID: 2154599
  47. In vitro transcriptional activation, dimerization, and DNA-binding specificity of the Epstein-Barr virus Zta protein.
    J Virol. 1990 Jun;64(6):2560-8 PMID: 2159531
  48. Analysis of the protein-coding content of the sequence of human cytomegalovirus strain AD169.
    Curr Top Microbiol Immunol. 1990;154:125-69 PMID: 2161319
  49. Two binding sites for the herpes simplex virus type 1 UL9 protein are required for efficient activity of the oriS replication origin.
    J Gen Virol. 1990 Jun;71 ( Pt 6):1379-85 PMID: 2161905
  50. The Epstein-Barr virus Zta transactivator: a member of the bZIP family with unique DNA-binding specificity and a dimerization domain that lacks the characteristic heptad leucine zipper motif.
    J Virol. 1990 Jul;64(7):3358-69 PMID: 2161945
  51. The Epstein-Barr virus (EBV) BMRF1 promoter for early antigen (EA-D) is regulated by the EBV transactivators, BRLF1 and BZLF1, in a cell-specific manner.
    J Virol. 1990 Aug;64(8):3753-9 PMID: 2164595
  52. Kaposi's sarcoma-associated herpesvirus-like DNA sequences in multicentric Castleman's disease.
    Blood. 1995 Aug 15;86(4):1276-80 PMID: 7632932
  53. In vitro establishment and characterization of two acquired immunodeficiency syndrome-related lymphoma cell lines (BC-1 and BC-2) containing Kaposi's sarcoma-associated herpesvirus-like (KSHV) DNA sequences.
    Blood. 1995 Oct 1;86(7):2708-14 PMID: 7670109
  54. Detection of Kaposi sarcoma associated herpesvirus in peripheral blood of HIV-infected individuals and progression to Kaposi's sarcoma.
    Lancet. 1995 Sep 23;346(8978):799-802 PMID: 7674745
  55. A general mechanism for transcriptional synergy by eukaryotic activators.
    Nature. 1995 Sep 21;377(6546):254-7 PMID: 7675113
  56. Adenovirus replication is coupled with the dynamic properties of the PML nuclear structure.
    Genes Dev. 1996 Jan 15;10(2):196-207 PMID: 8566753
  57. Comparing transcriptional activation and autostimulation by ZEBRA and ZEBRA/c-Fos chimeras.
    J Virol. 1996 Mar;70(3):1493-504 PMID: 8627667
  58. Functional order of assembly of herpes simplex virus DNA replication proteins into prereplicative site structures.
    J Virol. 1996 Mar;70(3):1759-67 PMID: 8627698
  59. Characterization of a binding site for the herpes simplex virus type 1 UL9 origin-binding protein within the UL9 gene.
    J Gen Virol. 1996 May;77 ( Pt 5):969-76 PMID: 8609494
  60. The periphery of nuclear domain 10 (ND10) as site of DNA virus deposition.
    J Cell Biol. 1996 Aug;134(4):815-26 PMID: 8769408
  61. Epstein-Barr viral latency is disrupted by the immediate-early BRLF1 protein through a cell-specific mechanism.
    Proc Natl Acad Sci U S A. 1996 Aug 20;93(17):9194-9 PMID: 8799177
  62. Evidence that the UL84 gene product of human cytomegalovirus is essential for promoting oriLyt-dependent DNA replication and formation of replication compartments in cotransfection assays.
    J Virol. 1996 Nov;70(11):7398-413 PMID: 8892858
  63. Kaposi's sarcoma-associated herpesvirus contains G protein-coupled receptor and cyclin D homologs which are expressed in Kaposi's sarcoma and malignant lymphoma.
    J Virol. 1996 Nov;70(11):8218-23 PMID: 8892957
  64. Nucleotide sequence of the Kaposi sarcoma-associated herpesvirus (HHV8).
    Proc Natl Acad Sci U S A. 1996 Dec 10;93(25):14862-7 PMID: 8962146
  65. A replication function associated with the activation domain of the Epstein-Barr virus Zta transactivator.
    J Virol. 1996 Dec;70(12):8340-7 PMID: 8970953
  66. Selective switch between latency and lytic replication of Kaposi's sarcoma herpesvirus and Epstein-Barr virus in dually infected body cavity lymphoma cells.
    J Virol. 1997 Jan;71(1):314-24 PMID: 8985352
  67. A single 13-kilobase divergent locus in the Kaposi sarcoma-associated herpesvirus (human herpesvirus 8) genome contains nine open reading frames that are homologous to or related to cellular proteins.
    J Virol. 1997 Mar;71(3):1963-74 PMID: 9032328
  68. Formation of herpes simplex virus type 1 replication compartments by transfection: requirements and localization to nuclear domain 10.
    J Virol. 1997 Mar;71(3):2390-9 PMID: 9032376
  69. Assembly of complete, functionally active herpes simplex virus DNA replication compartments and recruitment of associated viral and cellular proteins in transient cotransfection assays.
    J Virol. 1997 Apr;71(4):3146-60 PMID: 9060678
  70. Assembly of herpes simplex virus replication proteins at two distinct intranuclear sites.
    Virology. 1997 Mar 3;229(1):113-25 PMID: 9123852
  71. Herpes simplex virus type 1 prereplicative sites are a heterogeneous population: only a subset are likely to be precursors to replication compartments.
    J Virol. 1997 Jun;71(6):4771-81 PMID: 9151871
  72. Human cytomegalovirus immediate early interaction with host nuclear structures: definition of an immediate transcript environment.
    J Cell Biol. 1997 Jul 14;138(1):5-16 PMID: 9214377
  73. Export of importin alpha from the nucleus is mediated by a specific nuclear transport factor.
    Cell. 1997 Sep 19;90(6):1061-71 PMID: 9323134
  74. Identification and characterization of human herpesvirus-8 lytic cycle-associated ORF 59 protein and the encoding cDNA by monoclonal antibody.
    Virology. 1998 Jan 5;240(1):118-26 PMID: 9448696
  75. The Epstein-Barr virus lytic transactivator Zta interacts with the helicase-primase replication proteins.
    J Virol. 1998 Nov;72(11):8559-67 PMID: 9765394
  76. Mta has properties of an RNA export protein and increases cytoplasmic accumulation of Epstein-Barr virus replication gene mRNA.
    J Virol. 1998 Dec;72(12):9526-34 PMID: 9811685
  77. ND10 protein PML is recruited to herpes simplex virus type 1 prereplicative sites and replication compartments in the presence of viral DNA polymerase.
    J Virol. 1998 Dec;72(12):10100-7 PMID: 9811750
  78. Kaposi's sarcoma-associated herpesvirus encodes a bZIP protein with homology to BZLF1 of Epstein-Barr virus.
    J Virol. 1999 Mar;73(3):1909-17 PMID: 9971770
Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.75.3.1487-1506.2001
Published
2001-02-00
Pages
1487-506
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC114054
Subset
IM
Grants
NCI NIH HHS · R01 CA073585 · United States
NCI NIH HHS · T32 CA009243 · United States
NCI NIH HHS · R01 CA73585 · United States
NCI NIH HHS · T32 CA09243 · United States
External Links
PubMed source DOI: 10.1128/JVI.75.3.1487-1506.2001 PMC Full Text View journal details
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