Transcription factor NF-kappa B is activated by photosensitization generating oxidative DNA damages.
Reactive oxygen intermediates like hydrogen peroxide (H2O2) have been shown to serve as messengers in the induction of NF-kappa B and, then, in the activation and replication of human immunodeficiency virus (HIV)-1 in human cells. Because H2O2 can be converted into the highly reactive OH. at various locations inside the cells, we started to investigate the generation of Reactive oxygen intermediates by photosensitization. This technique is based on the use of a photosensitizer which is a molecule absorbing visible light and which can be located at various sites inside the cell depending on its physicochemical properties. In this work, we used proflavine (PF), a cationic molecule having a high affinity for DNA, capable of intercalating between DNA base pairs. Upon visible light irradiation, intercalated PF molecules oxidize guanine residues and generate DNA single-strand breaks. In lymphocytes or monocytes latently infected with HIV-1 (ACH-2 or U1, respectively), this photosensitizing treatment induced a cytotoxicity, an induction of NF-kappa B, and a reactivation of HIV-1 in cells surviving the treatment. NF-kappa B induction by PF-mediated photosensitization was not affected by the presence of N-acetyl-L-cysteine while strong inhibition was recorded when the induction was triggered by H2O2 or by phorbol 12-myristate 13-acetate. Another transcription factor like AP-1 is less activated by this photosensitizing treatment. In comparison with other inducing treatments, such as phorbol 12-myristate 13-acetate or tumor necrosis factor alpha, the activation of NF-kappa B is slow, being optimal 120 min after treatment. These kinetic data were obtained by following, on the same samples, both the appearance of NF-kappa B in the nucleus and the disappearance of I kappa B-alpha in cytoplasmic extracts. These data allow us to postulate that signaling events, initiated by DNA oxidative damages, are transmitted into the cytoplasm where the inactive NF-kappa B factor is resident and allow the translocation of p50/p65 subunits of NF-kappa B to the nucleus leading to HIV-1 gene expression.
Legrand-Poels
S
S
Laboratory of Virology, University of Liège, Belgium.
Bours
V
V
Piret
B
B
Pflaum
M
M
Epe
B
B
Rentier
B
B
Piette
J
J
D000111:Acetylcysteine / Q000494:pharmacology; D001483:Base Sequence; D004247:DNA / Q000378:metabolism; D004249:DNA Damage; D015497:HIV-1 / Q000502:physiology; D006861:Hydrogen Peroxide / Q000494:pharmacology; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000378:metabolism*; D018384:Oxidative Stress; D017319:Photosensitizing Agents / Q000494:pharmacology*; D011370:Proflavine / Q000494:pharmacology; D018808:Transcription Factor AP-1 / Q000378:metabolism; D014775:Virus Activation
D016328:NF-kappa B; D017319:Photosensitizing Agents; D018808:Transcription Factor AP-1; D004247:DNA; D006861:Hydrogen Peroxide; D011370:Proflavine; D000111:Acetylcysteine