Coinduction of c-jun gene expression and internucleosomal DNA fragmentation by ionizing radiation.
Previous work has demonstrated that the cellular response to ionizing radiation includes transcriptional activation of the c-jun early response gene. The present studies demonstrate that this induction of c-jun expression is temporally related to the appearance of internucleosomal DNA fragmentation. These events were maximal at 6 h and transient after exposure to lethal doses (20 Gy) of ionizing radiation. We also demonstrate that N-acetyl-L-cysteine (NAC), an antioxidant, inhibits X-ray-induced c-jun expression and endonucleolytic DNA cleavage. These findings suggested that both events are mediated at least in part through the formation of reactive oxygen intermediates (ROIs). Since ROIs damage DNA and X-ray-induced DNA damage is associated with activation of poly(ADP-ribose) polymerase (ADPRP), we studied the effects of the ADPRP inhibitors 3-aminobenzamide (3-AB), nicotinamide, and theophylline. 3-AB blocked both X-ray-induced c-jun expression and internucleosomal DNA fragmentation. Similar findings were obtained with nicotinamide and theophylline. In contrast, 3-AB had little if any effect on induction of c-jun transcripts or DNA fragmentation induced by the alkylating agent mitomycin C. While c-jun expression is restricted to cells in G1 and G1/S phases, we have found that X-ray-induced c-jun transcripts are detectable throughout all phases of the cell cycle. The induction of internucleosomal DNA fragmentation by X-rays was also detectable throughout the cell cycle. Taken together, these results support the coinduction of c-jun transcription and internucleosomal DNA fragmentation by ionizing radiation. Similar studies were performed with H2O2 since this agent also results in the production of ROIs.(ABSTRACT TRUNCATED AT 250 WORDS)
Laboratory of Clinical Pharmacology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts 02115.
Y
Y
Manome
R
R
Datta
N
N
Taneja
T
T
Shafman
E
E
Bump
R
R
Hass
R
R
Weichselbaum
D
D
Kufe
D002460:Cell Line; D004247:DNA / Q000187:drug effects / Q000302:isolation & purification / Q000528:radiation effects*; D004249:DNA Damage; D015870:Gene Expression / Q000187:drug effects / Q000235:genetics*; D016758:Genes, jun / Q000187:drug effects / Q000528:radiation effects*; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D009536:Niacinamide / Q000494:pharmacology; D009707:Nucleosomes / Q000378:metabolism / Q000528:radiation effects*; D016760:Proto-Oncogene Proteins c-fos / Q000096:biosynthesis; D016755:Proto-Oncogene Proteins c-jun / Q000096:biosynthesis; D013806:Theophylline / Q000494:pharmacology; D014965:X-Rays
D009707:Nucleosomes; D016760:Proto-Oncogene Proteins c-fos; D016755:Proto-Oncogene Proteins c-jun; D009536:Niacinamide; D004247:DNA; D006861:Hydrogen Peroxide; D013806:Theophylline