Home LiteratureArticle Details
PMID: 10196319 Published · ppublish English Journal Article

The amino-terminal region of Vpr from human immunodeficiency virus type 1 forms ion channels and kills neurons.

Journal of virology ·Vol. 73 ·No. 5 ·1999-05-00 ·Pages 4230-8

Piller SC, Ewart GD, Jans DA, Gage PW, Cox GB

Abstract

We have previously reported that the accessory protein Vpr from human immunodeficiency virus type 1 forms cation-selective ion channels in planar lipid bilayers and is able to depolarize intact cultured neurons by causing an inward sodium current, resulting in cell death. In this study, we used site-directed mutagenesis and synthetic peptides to identify the structural regions responsible for the above functions. Mutations in the N-terminal region of Vpr were found to affect channel activity, whereas this activity was not affected by mutations in the hydrophobic region of Vpr (amino acids 53 to 71). Analysis of mutants containing changes in the basic C terminus confirmed previous results that this region, although not necessary for ion channel function, was responsible for the observed rectification of wild-type Vpr currents. A peptide comprising the first 40 N-terminal amino acids of Vpr (N40) was found to be sufficient to form ion channels similar to those caused by wild-type Vpr in planar lipid bilayers. Furthermore, N40 was able to cause depolarization of the plasmalemma and cell death in cultured hippocampal neurons with a time course similar to that seen with wild-type Vpr, supporting the idea that this region is responsible for Vpr ion channel function and cytotoxic effects. Since Vpr is found in the serum and cerebrospinal fluids of AIDS patients, these results may have significance for AIDS pathology.

MeSH Terms
Amino Acid Sequence Animals Cell Death Gene Products, vpr/genetics,metabolism,physiology HIV-1/metabolism Humans Molecular Sequence Data Mutagenesis, Site-Directed Neurons/cytology,physiology Rats Sodium Channels/genetics,metabolism,physiology Structure-Activity Relationship vpr Gene Products, Human Immunodeficiency Virus
Chemicals
Gene Products, vpr Sodium Channels vpr Gene Products, Human Immunodeficiency Virus
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Piller S C
Membrane Biology Program, John Curtin School of Medical Research, Australian National University, Canberra, ACT 2601, Australia.
Ewart G D
Jans D A
Gage P W
Cox G B
References (40)
40 references, click to expand
  1. Human immunodeficiency virus type 1 viral protein R localization in infected cells and virions.
    J Virol. 1993 Nov;67(11):6542-50 PMID: 8411357
  2. HIV-1 protein Vpr causes gross mitochondrial dysfunction in the yeast Saccharomyces cerevisiae.
    FEBS Lett. 1997 Jun 30;410(2-3):145-9 PMID: 9237618
  3. Flow cytometric analysis of membrane potential in embryonic rat spinal cord cells.
    J Neurosci Methods. 1988 Jan;22(3):203-13 PMID: 3361946
  4. Vpr protein of human immunodeficiency virus type 1 forms cation-selective channels in planar lipid bilayers.
    Proc Natl Acad Sci U S A. 1996 Jan 9;93(1):111-5 PMID: 8552585
  5. Extracellular addition of a domain of HIV-1 Vpr containing the amino acid sequence motif H(S/F)RIG causes cell membrane permeabilization and death.
    Mol Microbiol. 1996 Mar;19(6):1185-92 PMID: 8730861
  6. 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
  7. 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
  8. HIV-1, Vpr and the cell cycle.
    Curr Biol. 1996 Sep 1;6(9):1096-103 PMID: 8805364
  9. 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
  10. 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
  11. Vpr-induced cell cycle arrest is conserved among primate lentiviruses.
    J Virol. 1996 Apr;70(4):2516-24 PMID: 8642681
  12. Structural studies of synthetic peptide fragments derived from the HIV-1 Vpr protein.
    Biochem Biophys Res Commun. 1998 Mar 27;244(3):732-6 PMID: 9535734
  13. Nuclear transport of granzyme B (fragmentin-2). Dependence of perforin in vivo and cytosolic factors in vitro.
    J Biol Chem. 1996 Nov 29;271(48):30781-9 PMID: 8940058
  14. Functional analysis of HIV-1 Vpr: identification of determinants essential for subcellular localization.
    Virology. 1995 Oct 1;212(2):331-9 PMID: 7571402
  15. Influenza A virus M2 ion channel protein: a structure-function analysis.
    J Virol. 1994 Mar;68(3):1551-63 PMID: 7508997
  16. The Vpu protein of human immunodeficiency virus type 1 forms cation-selective ion channels.
    J Virol. 1996 Oct;70(10):7108-15 PMID: 8794357
  17. Role of the karyopherin pathway in human immunodeficiency virus type 1 nuclear import.
    J Virol. 1996 Feb;70(2):1027-32 PMID: 8551560
  18. Coupled potassium channels induced by arachidonic acid in cultured neurons.
    Proc Biol Sci. 1990 Oct 22;242(1303):17-22 PMID: 1980738
  19. A functionally defined model for the M2 proton channel of influenza A virus suggests a mechanism for its ion selectivity.
    Proc Natl Acad Sci U S A. 1997 Oct 14;94(21):11301-6 PMID: 9326604
  20. Human immunodeficiency virus type 1 Vpr induces apoptosis following cell cycle arrest.
    J Virol. 1997 Jul;71(7):5579-92 PMID: 9188632
  21. 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
  22. Extracellular HIV-1 virus protein R causes a large inward current and cell death in cultured hippocampal neurons: implications for AIDS pathology.
    Proc Natl Acad Sci U S A. 1998 Apr 14;95(8):4595-600 PMID: 9539783
  23. Mutagenic analysis of human immunodeficiency virus type 1 Vpr: role of a predicted N-terminal alpha-helical structure in Vpr nuclear localization and virion incorporation.
    J Virol. 1995 Nov;69(11):7032-44 PMID: 7474123
  24. Evolution of the primate lentiviruses: evidence from vpx and vpr.
    EMBO J. 1992 Sep;11(9):3405-12 PMID: 1324171
  25. Arginine residues in the C-terminus of HIV-1 Vpr are important for nuclear localization and cell cycle arrest.
    Virology. 1998 Mar 15;242(2):414-24 PMID: 9514978
  26. Mutagenesis of the putative alpha-helical domain of the Vpr protein of human immunodeficiency virus type 1: effect on stability and virion incorporation.
    Proc Natl Acad Sci U S A. 1995 Apr 25;92(9):3794-8 PMID: 7731985
  27. Serum Vpr regulates productive infection and latency of human immunodeficiency virus type 1.
    Proc Natl Acad Sci U S A. 1994 Nov 8;91(23):10873-7 PMID: 7971975
  28. Membrane potentials in yeast cells measured by direct and indirect methods.
    Biochim Biophys Acta. 1981 Apr 22;643(1):265-8 PMID: 7016192
  29. Inhibition of prokaryotic cell growth by HIV1 Vpr.
    Res Virol. 1997 May-Jun;148(3):207-13 PMID: 9201811
  30. Distinct effects in primary macrophages and lymphocytes of the human immunodeficiency virus type 1 accessory genes vpr, vpu, and nef: mutational analysis of a primary HIV-1 isolate.
    Virology. 1994 May 1;200(2):623-31 PMID: 8178448
  31. 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
  32. A C-terminal domain of HIV-1 accessory protein Vpr is involved in penetration, mitochondrial dysfunction and apoptosis of human CD4+ lymphocytes.
    Apoptosis. 1997;2(1):69-76 PMID: 14646566
  33. The vpr regulatory gene of HIV.
    Curr Top Microbiol Immunol. 1995;193:209-36 PMID: 7648876
  34. Biochemical mechanism of HIV-1 Vpr function. Oligomerization mediated by the N-terminal domain.
    J Biol Chem. 1994 Dec 23;269(51):32131-7 PMID: 7798208
  35. Ion channels formed by NB, an influenza B virus protein.
    J Membr Biol. 1996 Mar;150(2):127-32 PMID: 8661776
  36. Identification of an ion channel activity of the Vpu transmembrane domain and its involvement in the regulation of virus release from HIV-1-infected cells.
    FEBS Lett. 1996 Nov 25;398(1):12-8 PMID: 8946945
  37. Measurement of the electrochemical proton gradient in submitochondrial particles.
    J Biol Chem. 1983 Feb 10;258(3):1474-86 PMID: 6296098
  38. Identification of HIV-1 vpr product and function.
    J Acquir Immune Defic Syndr. 1990;3(1):11-8 PMID: 2136707
  39. Interaction with phospholipid bilayers, ion channel formation, and antimicrobial activity of basic amphipathic alpha-helical model peptides of various chain lengths.
    J Biol Chem. 1991 Oct 25;266(30):20218-22 PMID: 1718959
  40. Identification of residues in the N-terminal acidic domain of HIV-1 Vpr essential for virion incorporation.
    Virology. 1995 Feb 20;207(1):297-302 PMID: 7871742
Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
Published
1999-05-00
Pages
4230-8
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC104202
Subset
IM
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