Home LiteratureArticle Details
PMID: 17626091 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

DDB1 and Cul4A are required for human immunodeficiency virus type 1 Vpr-induced G2 arrest.

Journal of virology ·Vol. 81 ·No. 19 ·2007-10-00 ·Pages 10822-30

Tan L, Ehrlich E, Yu XF

Abstract

Vpr-mediated induction of G2 cell cycle arrest has been postulated to be important for human immunodeficiency virus type 1 (HIV-1) replication, but the precise role of Vpr in this cell cycle arrest is unclear. In the present study, we have shown that HIV-1 Vpr interacts with damaged DNA binding protein 1 (DDB1) but not its partner DDB2. The interaction of Vpr with DDB1 was inhibited when DCAF1 (VprBP) expression was reduced by short interfering RNA (siRNA) treatment. The Vpr mutant (Q65R) that was defective for DCAF1 interaction also had a defect in DDB1 binding. However, Vpr binding to DDB1 was not sufficient to induce G2 arrest. A reduction in DDB1 or DDB2 expression in the absence of Vpr also did not induce G2 arrest. On the other hand, Vpr-induced G2 arrest was impaired when the intracellular level of DDB1 or Cullin 4A was reduced by siRNA treatment. Furthermore, Vpr-induced G2 arrest was largely abolished by a proteasome inhibitor. These data suggest that Vpr assembles with DDB1 through interaction with DCAF1 to form an E3 ubiquitin ligase that targets cellular substrates for proteasome-mediated degradation and G2 arrest.

MeSH Terms
Amino Acid Sequence Cell Line Cullin Proteins/antagonists & inhibitors,genetics,metabolism DNA-Binding Proteins/antagonists & inhibitors,genetics,metabolism G2 Phase Gene Products, vpr/antagonists & inhibitors,genetics,metabolism HIV-1/physiology Humans Molecular Sequence Data Mutation Proteasome Endopeptidase Complex/metabolism Proteasome Inhibitors RNA, Small Interfering/genetics,pharmacology Ubiquitin-Protein Ligases Virus Replication vpr Gene Products, Human Immunodeficiency Virus
Chemicals
CUL4A protein, human Cullin Proteins DDB1 protein, human DDB2 protein, human DNA-Binding Proteins Gene Products, vpr Proteasome Inhibitors RNA, Small Interfering vpr Gene Products, Human Immunodeficiency Virus Ubiquitin-Protein Ligases Proteasome Endopeptidase Complex
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Tan Lindi
Department of Molecular Microbiology and Immunology, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD 21205, USA.
Ehrlich Elana
Yu Xiao-Fang
References (66)
66 references, click to expand
  1. Isolation of a cDNA encoding a UV-damaged DNA binding factor defective in xeroderma pigmentosum group E cells.
    Mutat Res. 1996 Jan 2;362(1):105-17 PMID: 8538642
  2. Phosphorylation of a novel SOCS-box regulates assembly of the HIV-1 Vif-Cul5 complex that promotes APOBEC3G degradation.
    Genes Dev. 2004 Dec 1;18(23):2861-6 PMID: 15574592
  3. Mutational alteration of human immunodeficiency virus type 1 Vif allows for functional interaction with nonhuman primate APOBEC3G.
    J Virol. 2006 Jun;80(12):5984-91 PMID: 16731937
  4. Docking of HIV-1 Vpr to the nuclear envelope is mediated by the interaction with the nucleoporin hCG1.
    J Biol Chem. 2002 Nov 22;277(47):45091-8 PMID: 12228227
  5. Cullin 4A associates with the UV-damaged DNA-binding protein DDB.
    J Biol Chem. 1999 Dec 10;274(50):35309-12 PMID: 10585395
  6. Induction of APOBEC3G ubiquitination and degradation by an HIV-1 Vif-Cul5-SCF complex.
    Science. 2003 Nov 7;302(5647):1056-60 PMID: 14564014
  7. Human immunodeficiency virus type 1 Vpr induces the degradation of the UNG and SMUG uracil-DNA glycosylases.
    J Virol. 2005 Sep;79(17):10978-87 PMID: 16103149
  8. Human immunodeficiency virus type 1 viral protein R (Vpr) arrests cells in the G2 phase of the cell cycle by inhibiting p34cdc2 activity.
    J Virol. 1995 Nov;69(11):6705-11 PMID: 7474080
  9. Mutational analysis of cell cycle arrest, nuclear localization and virion packaging of human immunodeficiency virus type 1 Vpr.
    J Virol. 1995 Dec;69(12):7909-16 PMID: 7494303
  10. The paramyxovirus simian virus 5 V protein slows progression of the cell cycle.
    J Virol. 2000 Oct;74(19):9152-66 PMID: 10982362
  11. Hepatitis B virus X protein and simian virus 5 V protein exhibit similar UV-DDB1 binding properties to mediate distinct activities.
    J Virol. 2003 Jun;77(11):6274-83 PMID: 12743284
  12. Primate lentiviral virion infectivity factors are substrate receptors that assemble with cullin 5-E3 ligase through a HCCH motif to suppress APOBEC3G.
    Proc Natl Acad Sci U S A. 2005 Aug 9;102(32):11444-9 PMID: 16076960
  13. Viral infections and cell cycle G2/M regulation.
    Cell Res. 2005 Mar;15(3):143-9 PMID: 15780175
  14. Human immunodeficiency virus type 1 Vpr arrests the cell cycle in G2 by inhibiting the activation of p34cdc2-cyclin B.
    J Virol. 1995 Nov;69(11):6859-64 PMID: 7474100
  15. Phosphorylation of human immunodeficiency virus type 1 Vpr regulates cell cycle arrest.
    J Virol. 2000 Jul;74(14):6520-7 PMID: 10864665
  16. Phosphorylation of Vpr regulates HIV type 1 nuclear import and macrophage infection.
    AIDS Res Hum Retroviruses. 2002 Mar 1;18(4):283-8 PMID: 11860675
  17. HIV-1 Vpr interacts with the nuclear transport pathway to promote macrophage infection.
    Genes Dev. 1998 Jan 15;12(2):175-85 PMID: 9436978
  18. Human immunodeficiency virus type 1 cell cycle control: Vpr is cytostatic and mediates G2 accumulation by a mechanism which differs from DNA damage checkpoint control.
    J Virol. 1996 Apr;70(4):2324-31 PMID: 8642659
  19. A novel inducible expression system to study transdominant mutants of HIV-1 Vpr.
    Virology. 2001 Aug 15;287(1):133-42 PMID: 11504548
  20. Xeroderma pigmentosum complementation group E and UV-damaged DNA-binding protein.
    DNA Repair (Amst). 2002 Aug 6;1(8):601-16 PMID: 12509284
  21. Viral protein R regulates docking of the HIV-1 preintegration complex to the nuclear pore complex.
    J Biol Chem. 1998 May 22;273(21):13347-52 PMID: 9582382
  22. Progression to AIDS in the absence of a gene for vpr or vpx.
    J Virol. 1995 Apr;69(4):2378-83 PMID: 7884883
  23. Lentiviral delivery of HIV-1 Vpr protein induces apoptosis in transformed cells.
    Proc Natl Acad Sci U S A. 1999 Oct 12;96(21):12039-43 PMID: 10518572
  24. HIV1 Vpr arrests the cell cycle by recruiting DCAF1/VprBP, a receptor of the Cul4-DDB1 ubiquitin ligase.
    Cell Cycle. 2007 Jan 15;6(2):182-8 PMID: 17314515
  25. Role of the karyopherin pathway in human immunodeficiency virus type 1 nuclear import.
    J Virol. 1996 Feb;70(2):1027-32 PMID: 8551560
  26. Roles of p53 and caspases in the induction of cell cycle arrest and apoptosis by HIV-1 vpr.
    Exp Cell Res. 1999 Aug 25;251(1):156-65 PMID: 10438581
  27. Cul4A and DDB1 associate with Skp2 to target p27Kip1 for proteolysis involving the COP9 signalosome.
    Mol Cell Biol. 2006 Apr;26(7):2531-9 PMID: 16537899
  28. Natural variation in Vif: differential impact on APOBEC3G/3F and a potential role in HIV-1 diversification.
    PLoS Pathog. 2005 Sep;1(1):e6 PMID: 16201018
  29. Molecular architecture and assembly of the DDB1-CUL4A ubiquitin ligase machinery.
    Nature. 2006 Oct 5;443(7111):590-3 PMID: 16964240
  30. The interaction of vpr with uracil DNA glycosylase modulates the human immunodeficiency virus type 1 In vivo mutation rate.
    J Virol. 2000 Aug;74(15):7039-47 PMID: 10888643
  31. Selective assembly of HIV-1 Vif-Cul5-ElonginB-ElonginC E3 ubiquitin ligase complex through a novel SOCS box and upstream cysteines.
    Genes Dev. 2004 Dec 1;18(23):2867-72 PMID: 15574593
  32. Viral protein R regulates nuclear import of the HIV-1 pre-integration complex.
    EMBO J. 1998 Feb 16;17(4):909-17 PMID: 9463369
  33. Roles of uracil-DNA glycosylase and dUTPase in virus replication.
    J Gen Virol. 2002 Oct;83(Pt 10):2339-45 PMID: 12237414
  34. Relationship of the xeroderma pigmentosum group E DNA repair defect to the chromatin and DNA binding proteins UV-DDB and replication protein A.
    Mol Cell Biol. 1998 Jun;18(6):3182-90 PMID: 9584159
  35. Two E3 ubiquitin ligases, SCF-Skp2 and DDB1-Cul4, target human Cdt1 for proteolysis.
    EMBO J. 2006 Mar 8;25(5):1126-36 PMID: 16482215
  36. Human immunodeficiency virus type 1 Vpr induces apoptosis following cell cycle arrest.
    J Virol. 1997 Jul;71(7):5579-92 PMID: 9188632
  37. Interaction of the human immunodeficiency virus type 1 Vpr protein with the nuclear pore complex.
    J Virol. 1998 Jul;72(7):6004-13 PMID: 9621063
  38. HIV-1 Vpr increases viral expression by manipulation of the cell cycle: a mechanism for selection of Vpr in vivo.
    Nat Med. 1998 Jan;4(1):65-71 PMID: 9427608
  39. Targeted ubiquitination of CDT1 by the DDB1-CUL4A-ROC1 ligase in response to DNA damage.
    Nat Cell Biol. 2004 Oct;6(10):1003-9 PMID: 15448697
  40. DDB1 maintains genome integrity through regulation of Cdt1.
    Mol Cell Biol. 2006 Nov;26(21):7977-90 PMID: 16940174
  41. Open reading frame vpr of simian immunodeficiency virus encodes a virion-associated protein.
    J Virol. 1990 Nov;64(11):5688-93 PMID: 2145446
  42. The human immunodeficiency virus type 1 vpr gene arrests infected T cells in the G2 + M phase of the cell cycle.
    J Virol. 1995 Oct;69(10):6304-13 PMID: 7666531
  43. Comparison of cell cycle arrest, transactivation, and apoptosis induced by the simian immunodeficiency virus SIVagm and human immunodeficiency virus type 1 vpr genes.
    J Virol. 2001 Apr;75(8):3791-801 PMID: 11264368
  44. HIV-1 Vpr function is mediated by interaction with the damage-specific DNA-binding protein DDB1.
    Proc Natl Acad Sci U S A. 2007 Mar 6;104(10):4130-5 PMID: 17360488
  45. HIV-1 Vpr potently induces programmed cell death in the CNS in vivo.
    DNA Cell Biol. 2007 Feb;26(2):116-31 PMID: 17328670
  46. HIV-1 Vpr activates the G2 checkpoint through manipulation of the ubiquitin proteasome system.
    Virol J. 2007;4:57 PMID: 17559673
  47. Hepatitis B virus X protein associated with UV-DDB1 induces cell death in the nucleus and is functionally antagonized by UV-DDB2.
    J Biol Chem. 2002 Oct 11;277(41):38847-54 PMID: 12151405
  48. Vpr is required for efficient replication of human immunodeficiency virus type-1 in mononuclear phagocytes.
    Virology. 1995 Feb 1;206(2):935-44 PMID: 7531918
  49. The HIV-1 viral protein R induces apoptosis via a direct effect on the mitochondrial permeability transition pore.
    J Exp Med. 2000 Jan 3;191(1):33-46 PMID: 10620603
  50. DNA damage binding protein component DDB1 participates in nucleotide excision repair through DDB2 DNA-binding and cullin 4A ubiquitin ligase activity.
    Cancer Res. 2006 Sep 1;66(17):8590-7 PMID: 16951172
  51. Differential requirement for conserved tryptophans in human immunodeficiency virus type 1 Vif for the selective suppression of APOBEC3G and APOBEC3F.
    J Virol. 2006 Mar;80(6):3112-5 PMID: 16501124
  52. Nuclear transport of human DDB protein induced by ultraviolet light.
    J Biol Chem. 2000 Jul 14;275(28):21429-34 PMID: 10777491
  53. HIV-1 Vpr induces apoptosis through caspase 9 in T cells and peripheral blood mononuclear cells.
    J Biol Chem. 2002 Oct 4;277(40):37820-31 PMID: 12095993
  54. HIV-1 Vif protein binds the editing enzyme APOBEC3G and induces its degradation.
    Nat Med. 2003 Nov;9(11):1398-403 PMID: 14528301
  55. Human immunodeficiency virus vpr gene encodes a virion-associated protein.
    AIDS Res Hum Retroviruses. 1990 Nov;6(11):1265-71 PMID: 2150318
  56. DDB accumulates at DNA damage sites immediately after UV irradiation and directly stimulates nucleotide excision repair.
    J Biol Chem. 2002 Jan 18;277(3):1637-40 PMID: 11705987
  57. Cul4A targets p27 for degradation and regulates proliferation, cell cycle exit, and differentiation during erythropoiesis.
    Blood. 2006 Jun 1;107(11):4291-9 PMID: 16467204
  58. The role of Vpr in HIV-1 pathogenesis.
    Curr HIV Res. 2005 Jan;3(1):43-51 PMID: 15638722
  59. The Vpr protein from HIV-1: distinct roles along the viral life cycle.
    Retrovirology. 2005;2:11 PMID: 15725353
  60. The Vpr protein of human immunodeficiency virus type 1 influences nuclear localization of viral nucleic acids in nondividing host cells.
    Proc Natl Acad Sci U S A. 1994 Jul 19;91(15):7311-5 PMID: 8041786
  61. DDB complexities.
    DNA Repair (Amst). 2003 Sep 18;2(9):1065-9 PMID: 12967661
  62. Effect of HIV-1 Vpr on cell cycle regulators.
    DNA Cell Biol. 2004 Apr;23(4):249-60 PMID: 15142382
  63. DNA damage induces Cdt1 proteolysis in fission yeast through a pathway dependent on Cdt2 and Ddb1.
    EMBO Rep. 2006 Nov;7(11):1134-9 PMID: 17039252
  64. The Caenorhabditis elegans replication licensing factor CDT-1 is targeted for degradation by the CUL-4/DDB-1 complex.
    Mol Cell Biol. 2007 Feb;27(4):1394-406 PMID: 17145765
  65. Identification of HIV-1 vpr product and function.
    J Acquir Immune Defic Syndr. 1990;3(1):11-8 PMID: 2136707
  66. Vpr-mediated incorporation of UNG2 into HIV-1 particles is required to modulate the virus mutation rate and for replication in macrophages.
    J Biol Chem. 2004 Jul 2;279(27):28419-25 PMID: 15096517
Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.01380-07
Published
2007-10-00
Epub
2007-00-11
Pages
10822-30
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC2045451
Subset
IM
Grants
NIAID NIH HHS · R01 AI062644 · United States
NIAID NIH HHS · R01 AI062644-03 · United States
NIAID NIH HHS · R56 AI062644 · United States
NIAID NIH HHS · AI062644 · United States
External Links
PubMed source DOI: 10.1128/JVI.01380-07 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2