Home LiteratureArticle Details
PMID: 10846089 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

A carboxy-terminally truncated form of the human immunodeficiency virus type 1 Vpr protein induces apoptosis via G(1) cell cycle arrest.

Journal of virology ·Vol. 74 ·No. 13 ·2000-07-00 ·Pages 6058-67

Nishizawa M, Kamata M, Katsumata R, Aida Y

Abstract

Viral protein R (Vpr) of human immunodeficiency virus type 1 inhibits cell proliferation by arresting the cell cycle at the G(2) phase and inducing to apoptosis after G(2) arrest. We have reported previously that C81, a carboxy-terminally truncated form of Vpr, interferes with cell proliferation via a novel pathway that is distinct from G(2) arrest. However, the mechanism of this effect of C81 is unknown. We demonstrate here that C81 can induce apoptosis via G(1) arrest of the cell cycle. Immunostaining for various markers of stages of the cell cycle and flow cytometry analysis of DNA content showed that most HeLa cells that had been transiently transfected with a C81 expression vector were arrested at the G(1) phase and not at the G(2) or S phase of the cell cycle. Staining for annexin V, which binds phosphatidylserine on the plasma membrane, as an early indicator of apoptosis and measurement of the activity of caspase-3, a signaling molecule in apoptotic pathways, indicated that C81 is a strong inducer of apoptosis. Expression of C81 induced the condensation, fragmentation, and clumping of chromatin that are typical of apoptosis. Furthermore, the kinetics of the C81-induced G(1) arrest were closely correlated with changes in the number of annexin V-positive cells and the activity of caspase-3. Replacement of Ile or Leu residues by Pro at positions 60, 67, 74, and 81 within the leucine zipper-like domain of C81 revealed that Ile60, Leu67, and Ile74 play important roles both in the C81-induced G(1) arrest and in apoptosis. Thus, it appears that C81 induces apoptosis through pathways that are identical to those utilized for G(1) arrest of the cell cycle. It has been reported that Ile60, Leu67, and Ile74 also play an important role in the C81-induced suppression of growth. These results suggest that the suppression of growth induced by C81 result in apoptosis that is independent of G(2) arrest of the cell cycle.

MeSH Terms
Apoptosis G1 Phase Gene Products, vpr/genetics,metabolism HIV-1/genetics,metabolism HeLa Cells Humans Isoleucine/genetics,metabolism Leucine/genetics,metabolism Leucine Zippers Sequence Deletion vpr Gene Products, Human Immunodeficiency Virus
Chemicals
Gene Products, vpr vpr Gene Products, Human Immunodeficiency Virus Isoleucine Leucine
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Nishizawa M
Tsukuba Life Science Center, The Institute of Physical and Chemical Research (RIKEN), Ibaraki 305-0074, Japan.
Kamata M
Katsumata R
Aida Y
References (65)
65 references, click to expand
  1. Human immunodeficiency virus type 1 Vpr gene product prevents cell proliferation on mouse NIH3T3 cells without the G2 arrest of the cell cycle.
    Biochem Biophys Res Commun. 1997 Mar 17;232(2):550-4 PMID: 9125220
  2. p53 and ATM: cell cycle, cell death, and cancer.
    Adv Cancer Res. 1997;71:1-25 PMID: 9111862
  3. Human immunodeficiency virus type 1 Vpr induces apoptosis following cell cycle arrest.
    J Virol. 1997 Jul;71(7):5579-92 PMID: 9188632
  4. Death receptors: signaling and modulation.
    Science. 1998 Aug 28;281(5381):1305-8 PMID: 9721089
  5. Nef harbors a major determinant of pathogenicity for an AIDS-like disease induced by HIV-1 in transgenic mice.
    Cell. 1998 Oct 16;95(2):163-75 PMID: 9790524
  6. Characterization of HIV-1 vpr nuclear import: analysis of signals and pathways.
    J Cell Biol. 1998 Nov 16;143(4):875-85 PMID: 9817747
  7. Vpu increases susceptibility of human immunodeficiency virus type 1-infected cells to fas killing.
    J Virol. 1999 Jan;73(1):92-100 PMID: 9847311
  8. Human immunodeficiency virus type 1 Nef protein sensitizes CD4(+) T lymphoid cells to apoptosis via functional upregulation of the CD95/CD95 ligand pathway.
    Blood. 1999 Feb 1;93(3):1000-10 PMID: 9920849
  9. Requirements for efficient production and transduction of human immunodeficiency virus type 1-based vectors.
    J Virol. 1999 Mar;73(3):1828-34 PMID: 9971760
  10. Human immunodeficiency virus type 1 Tat induces apoptosis and increases sensitivity to apoptotic signals by up-regulating FLICE/caspase-8.
    J Virol. 1999 Mar;73(3):1956-63 PMID: 9971775
  11. Incorporation of Vpr into human immunodeficiency virus type 1 requires a direct interaction with the p6 domain of the p55 gag precursor.
    J Biol Chem. 1999 Mar 26;274(13):9083-91 PMID: 10085158
  12. Inhibition of prokaryotic cell growth by HIV1 Vpr.
    Res Virol. 1997 May-Jun;148(3):207-13 PMID: 9201811
  13. Nuclear import, virion incorporation, and cell cycle arrest/differentiation are mediated by distinct functional domains of human immunodeficiency virus type 1 Vpr.
    J Virol. 1997 Sep;71(9):6339-47 PMID: 9261351
  14. Early detection of apoptosis using a fluorescent conjugate of annexin V.
    Biotechniques. 1997 Sep;23(3):525-31 PMID: 9298227
  15. HIV-1 Vpr suppresses immune activation and apoptosis through regulation of nuclear factor kappa B.
    Nat Med. 1997 Oct;3(10):1117-23 PMID: 9334723
  16. Caspases: the executioners of apoptosis.
    Biochem J. 1997 Aug 15;326 ( Pt 1):1-16 PMID: 9337844
  17. Human immunodeficiency virus type 1 Vpr interacts with HHR23A, a cellular protein implicated in nucleotide excision DNA repair.
    J Virol. 1997 Dec;71(12):9732-42 PMID: 9371639
  18. Distinct mechanisms trigger apoptosis in human immunodeficiency virus type 1-infected and in uninfected bystander T lymphocytes.
    J Virol. 1998 Jan;72(1):660-70 PMID: 9420271
  19. 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
  20. HIV-1 Vpr interacts with the nuclear transport pathway to promote macrophage infection.
    Genes Dev. 1998 Jan 15;12(2):175-85 PMID: 9436978
  21. The HIV-1 vpr protein acts as a negative regulator of apoptosis in a human lymphoblastoid T cell line: possible implications for the pathogenesis of AIDS.
    J Exp Med. 1998 Feb 2;187(3):403-13 PMID: 9449720
  22. Viral protein R regulates nuclear import of the HIV-1 pre-integration complex.
    EMBO J. 1998 Feb 16;17(4):909-17 PMID: 9463369
  23. HIV-1 Vpr interacts with a human 34-kDa mov34 homologue, a cellular factor linked to the G2/M phase transition of the mammalian cell cycle.
    Proc Natl Acad Sci U S A. 1998 Mar 31;95(7):3419-24 PMID: 9520381
  24. HIV transcriptional activation by the accessory protein, VPR, is mediated by the p300 co-activator.
    Proc Natl Acad Sci U S A. 1998 Apr 28;95(9):5281-6 PMID: 9560267
  25. Cleavage of p21Cip1/Waf1 and p27Kip1 mediates apoptosis in endothelial cells through activation of Cdk2: role of a caspase cascade.
    Mol Cell. 1998 Mar;1(4):553-63 PMID: 9660939
  26. Cooperative actions of HIV-1 Vpr and p53 modulate viral gene transcription.
    J Biol Chem. 1998 Aug 7;273(32):20052-7 PMID: 9685344
  27. The HIV-1 Vpr displays strong anti-apoptotic activity.
    FEBS Lett. 1998 Jul 31;432(1-2):17-20 PMID: 9710242
  28. A carboxy-terminally truncated form of the Vpr protein of human immunodeficiency virus type 1 retards cell proliferation independently of G(2) arrest of the cell cycle.
    Virology. 1999 Oct 25;263(2):313-22 PMID: 10544105
  29. A role for the nuclear envelope in controlling DNA replication within the cell cycle.
    Nature. 1988 Apr 7;332(6164):546-8 PMID: 3357511
  30. Human immunodeficiency virus vpr product is a virion-associated regulatory protein.
    J Virol. 1990 Jun;64(6):3097-9 PMID: 2139896
  31. A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry.
    J Immunol Methods. 1991 Jun 3;139(2):271-9 PMID: 1710634
  32. Programmed death of T cells in HIV-1 infection.
    Science. 1992 Jul 10;257(5067):217-9 PMID: 1352911
  33. Induction of cell differentiation by human immunodeficiency virus 1 vpr.
    Cell. 1993 Feb 26;72(4):541-50 PMID: 8440020
  34. Direct and indirect mechanisms mediating apoptosis during HIV infection: contribution to in vivo CD4 T cell depletion.
    Semin Immunol. 1993 Jun;5(3):187-94 PMID: 8102263
  35. Human immunodeficiency virus type 1 viral protein R localization in infected cells and virions.
    J Virol. 1993 Nov;67(11):6542-50 PMID: 8411357
  36. Apoptosis induced in CD4+ cells expressing gp160 of human immunodeficiency virus type 1.
    J Virol. 1994 Jan;68(1):390-9 PMID: 7504739
  37. Requirement of the Pr55gag precursor for incorporation of the Vpr product into human immunodeficiency virus type 1 viral particles.
    J Virol. 1994 Mar;68(3):1926-34 PMID: 8107252
  38. Dicing with death: dissecting the components of the apoptosis machinery.
    Trends Biochem Sci. 1994 Jan;19(1):26-30 PMID: 8140617
  39. Activation of cyclin A-dependent protein kinases during apoptosis.
    Proc Natl Acad Sci U S A. 1994 Apr 26;91(9):3754-8 PMID: 8170983
  40. Biochemical mechanism of HIV-I Vpr function. Specific interaction with a cellular protein.
    J Biol Chem. 1994 Jun 3;269(22):15577-82 PMID: 8195203
  41. 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
  42. DNA polymerase alpha associated protein P1, a murine homolog of yeast MCM3, changes its intranuclear distribution during the DNA synthetic period.
    EMBO J. 1994 Sep 15;13(18):4311-20 PMID: 7925275
  43. Programmed cell death and AIDS: significance of T-cell apoptosis in pathogenic and nonpathogenic primate lentiviral infections.
    Proc Natl Acad Sci U S A. 1994 Sep 27;91(20):9431-5 PMID: 7937784
  44. Indirect mechanisms of HIV pathogenesis: how does HIV kill T cells?
    Curr Opin Immunol. 1994 Aug;6(4):605-15 PMID: 7946050
  45. The human immunodeficiency virus type 1 vpr gene prevents cell proliferation during chronic infection.
    J Virol. 1995 Feb;69(2):882-8 PMID: 7815556
  46. The p6gag domain of human immunodeficiency virus type 1 is sufficient for the incorporation of Vpr into heterologous viral particles.
    J Virol. 1995 May;69(5):2759-64 PMID: 7707498
  47. The glucocorticoid receptor type II complex is a target of the HIV-1 vpr gene product.
    Proc Natl Acad Sci U S A. 1995 Apr 11;92(8):3621-5 PMID: 7724608
  48. Programmed cell death and AIDS pathogenesis: significance and potential mechanisms.
    Curr Top Microbiol Immunol. 1995;200:195-211 PMID: 7634833
  49. 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
  50. Rapid induction of apoptosis by cell-to-cell transmission of human immunodeficiency virus type 1.
    J Virol. 1995 Oct;69(10):6457-65 PMID: 7666547
  51. Cell cycle and apoptosis: common pathways to life and death.
    J Cell Biochem. 1995 Jun;58(2):175-80 PMID: 7673325
  52. 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
  53. 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
  54. A leucine triplet repeat sequence (LXX)4 in p6gag is important for Vpr incorporation into human immunodeficiency virus type 1 particles.
    J Virol. 1995 Nov;69(11):6873-9 PMID: 7474102
  55. Interaction of virion protein Vpr of human immunodeficiency virus type 1 with cellular transcription factor Sp1 and trans-activation of viral long terminal repeat.
    J Biol Chem. 1995 Oct 27;270(43):25564-9 PMID: 7592727
  56. Apoptosis occurs predominantly in bystander cells and not in productively infected cells of HIV- and SIV-infected lymph nodes.
    Nat Med. 1995 Feb;1(2):129-34 PMID: 7585008
  57. A conserved LXXLF sequence is the major determinant in p6gag required for the incorporation of human immunodeficiency virus type 1 Vpr.
    J Virol. 1996 Jan;70(1):159-64 PMID: 8523520
  58. Human immunodeficiency virus type 1 Vpr protein binds to the uracil DNA glycosylase DNA repair enzyme.
    J Virol. 1996 Feb;70(2):697-704 PMID: 8551605
  59. Role of the karyopherin pathway in human immunodeficiency virus type 1 nuclear import.
    J Virol. 1996 Feb;70(2):1027-32 PMID: 8551560
  60. 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
  61. Vpr-induced cell cycle arrest is conserved among primate lentiviruses.
    J Virol. 1996 Apr;70(4):2516-24 PMID: 8642681
  62. Suppression of apoptosis by dominant negative mutants of cyclin-dependent protein kinases.
    J Biol Chem. 1996 Apr 26;271(17):10205-9 PMID: 8626584
  63. FACS-optimized mutants of the green fluorescent protein (GFP).
    Gene. 1996;173(1 Spec No):33-8 PMID: 8707053
  64. The human immunodeficiency virus type 1 Vpr transactivator: cooperation with promoter-bound activator domains and binding to TFIIB.
    J Mol Biol. 1996 Sep 6;261(5):599-606 PMID: 8800208
  65. Regulation of apoptosis by viral gene products.
    J Virol. 1997 Mar;71(3):1739-46 PMID: 9032302
Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/jvi.74.13.6058-6067.2000
Published
2000-07-00
Pages
6058-67
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC112104
Subset
IM
External Links
PubMed source DOI: 10.1128/jvi.74.13.6058-6067.2000 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