title: UV-induced apoptosis in XPG-deficient fibroblasts involves activation of CD95 and caspases but not p53.
abstract: Mildly affected individuals from xeroderma pigmentosum complementation group G (XP-G) possess single amino acid substitutions in the XPG protein that adversely affects its 3' endonuclease function in nucleotide excision repair. More serious mutations in the XPG gene generate truncated or unstable XPG proteins and result in a particularly early and severe form of the combined XP/CS complex. Following UV irradiation, cells from such XP-G/CS patients enter apoptosis more readily than other DNA repair-deficient cells. Here, we explore the mechanisms by which UV triggers the apoptotic cell death program in XP-G and XP-G/CS primary fibroblasts. Activation of the CD95 signalling pathway occurs within minutes and it is the earliest detectable post-UV event in such cells. This is rapidly followed by activation of caspase-8 then caspase-3. Several hours later caspase-9 becomes activated and the mitochondrial membrane potential drops, but without any obvious prior release of cytochrome c. Although p53 accumulates in XPG-deficient cells after UV irradiation, use of RNA interference demonstrates that p53 is not required for their UV-induced apoptotic response. p53 ablation of wild-type fibroblasts reduces MDM2 mRNA levels, inhibits accumulation of the 90kDa/92kDa Mdm2 isoforms, and prevents the nuclear relocalisation of Mdm2 after UV treatment. The same post-UV effects occur in XPG-deficient cells that express normal p53 levels. These results emphasise the importance of the extrinsic apoptotic pathway and aberrant Mdm2 events for the severe UV-induced apoptosis of XPG-deficient primary fibroblasts. XP-G/CS cells constitutively overexpress the pro-apoptotic Bax protein and a long isoform of the E2F1 transcription factor that controls S phase entry, which may prime them to enter apoptosis very readily after UV irradiation.
authors.affiliation: Department of Microbiology and Molecular Medicine, University Medical Centre (CMU), 1 rue Michel-Servet, 1211 Geneva 4, Switzerland. virginie.clement@medecine.unige.ch
authors.forename: Virginie
authors.initials: V
authors.lastname: Clément
authors.forename: Isabelle
authors.initials: I
authors.lastname: Dunand-Sauthier
authors.forename: Maciej
authors.initials: M
authors.lastname: Wiznerowicz
authors.forename: Stuart G
authors.initials: SG
authors.lastname: Clarkson
mesh_terms: D017209:Apoptosis / Q000528:radiation effects*; D053148:Caspase 3 / Q000378:metabolism*; D053453:Caspase 9 / Q000378:metabolism*; D002478:Cells, Cultured; D004260:DNA Repair; D004268:DNA-Binding Proteins / Q000235:genetics / Q000502:physiology*; D004720:Endonucleases / Q000235:genetics / Q000502:physiology*; D005347:Fibroblasts / Q000502:physiology / Q000528:radiation effects; D006801:Humans; D053078:Membrane Potential, Mitochondrial; D009687:Nuclear Proteins / Q000235:genetics / Q000502:physiology*; D051736:Proto-Oncogene Proteins c-mdm2 / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000502:physiology*; D014158:Transcription, Genetic; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism*; D014466:Ultraviolet Rays; D019014:fas Receptor / Q000378:metabolism*; D000099250:DNA Excision Repair Protein ERCC-5
chemical_list: D053148:Caspase 3; D053453:Caspase 9; D004268:DNA-Binding Proteins; D004720:Endonucleases; D009687:Nuclear Proteins; D051736:Proto-Oncogene Proteins c-mdm2; D014157:Transcription Factors; D016159:Tumor Suppressor Protein p53; D019014:fas Receptor; D000099250:DNA Excision Repair Protein ERCC-5; C497206:MDM2 protein, human