The amino-terminal region of Vpr from human immunodeficiency virus type 1 forms ion channels and kills neurons.
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.
Piller
S C
SC
Membrane Biology Program, John Curtin School of Medical Research, Australian National University, Canberra, ACT 2601, Australia.
Ewart
G D
GD
Jans
D A
DA
Gage
P W
PW
Cox
G B
GB
D000595:Amino Acid Sequence; D000818:Animals; D016923:Cell Death; D016336:Gene Products, vpr / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D015497:HIV-1 / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009474:Neurons / Q000166:cytology* / Q000502:physiology; D051381:Rats; D015222:Sodium Channels / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D013329:Structure-Activity Relationship; D054325:vpr Gene Products, Human Immunodeficiency Virus
D016336:Gene Products, vpr; D015222:Sodium Channels; D054325:vpr Gene Products, Human Immunodeficiency Virus