Human immunodeficiency virus type 1 vpr protein transactivation function: mechanism and identification of domains involved.
The human immunodeficiency virus type 1 (HIV-1) Vpr protein is a virion-associated protein that localizes in the nucleus of infected cells. Vpr has been shown to facilitate HIV infection of non-dividing cells such as macrophages by contributing to the nuclear translocation of the pre-integration complex. More recently, Vpr expression has been shown to induce an accumulation of cells at the G2 phase of the cell-cycle. We have previously reported that Vpr stimulates reporter gene expression directed from the HIV-1 long terminal repeat (LTR) as well as from heterologous viral promoters. However, the mode of action of Vpr-mediated transactivation remains to be precisely defined. We report here that, for a constant amount of transfected DNA, the level of chloramphenicol acetyltransferase (CAT) mRNA is increased in Vpr-expressing cells using either HIV-1 or a murine leukemia virus (MLV) SL3-3 LTR-CAT reporter construct. Moreover, this Vpr-mediated transactivation requires that promoters direct a minimal level of basal expression. Our mutagenic analysis indicates that the transactivation mediated by Vpr is not dependent on the ability of the protein to localize in the nucleus or to be packaged in the virions. Interestingly, all transactivation-competent Vpr mutants were still able to induce a cell-cycle arrest. Conversely, transactivation-defective mutants lost the ability to mediate cell-cycle arrest, implying a functional relationship between these two functions. Overall, our results indicate that the G2 cell-cycle arrest mediated by Vpr creates a cellular environment where the HIV-1 LTR is transcriptionally more active.
Forget
J
J
Département de Microbiologie et Immunologie, Université de Montréal, Montréal, Québec, H3C 3J7, Canada.
Yao
X J
XJ
Mercier
J
J
Cohen
E A
EA
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D016195:G2 Phase; D016336:Gene Products, vpr / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D017930:Genes, Reporter; D005814:Genes, Viral; D016325:HIV Long Terminal Repeat; D015497:HIV-1 / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D019169:Jurkat Cells; D009052:Leukemia Virus, Murine / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D010641:Phenotype; D015533:Transcriptional Activation; D054325:vpr Gene Products, Human Immunodeficiency Virus
D017931:DNA Primers; D004279:DNA, Viral; D016336:Gene Products, vpr; D054325:vpr Gene Products, Human Immunodeficiency Virus; D015500:Chloramphenicol O-Acetyltransferase