N-acetyl-L-cysteine-induced up-regulation of HIV-1 gene expression in monocyte-derived macrophages correlates with increased NF-kappaB DNA binding activity.
Nuclear factor kappaB (NF-kappaB) is an important cellular regulator of human immunodeficiency virus (HIV) gene expression. In T cells, N-acetyl-L-cysteine (NAC) inhibits the induction of NF-kappaB and transcription of HIV-1. However, NAC up-regulates HIV-1 replication in monocyte-derived macrophages (MDM). In this study we demonstrate that NAC treatment of MDM transfected with a chloramphenicol acetyltransferase (CAT) construct under transcriptional control of the HIV-1 long terminal repeat resulted in an up-regulation of CAT activity. Furthermore, MDM transfected with a HIV-1-NF-kappaB-CAT construct also produced increased CAT activity after NAC treatment. In addition, electrophoretic mobility shift assays revealed that nuclei of NAC-treated MDM contained increased binding activity to wild-type, but not mutant, kappaB oligonucleotides. Components of the binding activity were identified with antibodies as the NF-kappaB subunits p50 and p65. These data indicate that NAC-induced enhancement of HIV-1 replication in MDM is regulated at the level of viral gene expression and mediated by NF-kappaB.
Nottet
H S
HS
Eijkman-Winkler Institute of Medical Microbiology, University of Utrecht, The Netherlands.
Moelans
I I
II
de Vos
N M
NM
de Graaf
L
L
Visser
M R
MR
Verhoef
J
J
D000111:Acetylcysteine / Q000494:pharmacology*; D019380:Anti-HIV Agents / Q000494:pharmacology*; D002478:Cells, Cultured; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D004279:DNA, Viral / Q000378:metabolism*; D017930:Genes, Reporter / Q000187:drug effects; D005822:Genetic Vectors; D016325:HIV Long Terminal Repeat / Q000187:drug effects / Q000235:genetics; D015497:HIV-1 / Q000187:drug effects* / Q000235:genetics / Q000502:physiology; D006801:Humans; D019169:Jurkat Cells; D008264:Macrophages / Q000187:drug effects* / Q000378:metabolism / Q000821:virology; D016328:NF-kappa B / Q000378:metabolism*; D014158:Transcription, Genetic / Q000187:drug effects; D014162:Transfection; D015854:Up-Regulation / Q000187:drug effects; D014779:Virus Replication / Q000187:drug effects*
D019380:Anti-HIV Agents; D004279:DNA, Viral; D016328:NF-kappa B; D015500:Chloramphenicol O-Acetyltransferase; D000111:Acetylcysteine