Telomere attrition and genomic instability in xeroderma pigmentosum type-b deficient fibroblasts under oxidative stress.
Xeroderma pigmentosum B (XPB/ERCC3/p89) is an ATP-dependent 3'-->5' directed DNA helicase involved in basal RNA transcription and the nucleotide excision repair (NER) pathway. While the role of NER in alleviating oxidative DNA damage has been acknowledged it remains poorly understood. To study the involvement of XPB in repair of oxidative DNA damage, we utilized primary fibroblasts from a patient suffering from XP with Cockayne syndrome and hydrogen peroxide (H(2)O(2)) to induce oxidative stress. Mutant cells retained higher viability and cell cycle dysfunction after H(2)O(2) exposure. Cytokinesis blocked micronucleus assay revealed increased genome instability induced by H(2)O(2). Single cell gel electrophoresis (comet) assay showed that the missense mutation caused a reduced repair capacity for oxidative DNA damage. Mutant fibroblasts also displayed decreased population doubling rate, increased telomere attrition rate and early emergence of senescent characteristics under chronic low dose exposure to H(2)O(2). Fibroblasts from a heterozygous individual displayed intermediate traits in some assays and normal traits in others, indicating possible copy number dependence. The results show that a deficiency in functional XPB paradoxically renders cells more sensitive to the genotoxic effects of oxidative stress while reducing the cytotoxic effects. These findings have implications in the mechanisms of DNA repair, mutagenesis and carcinogenesis and ageing in normal physiological systems.
Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.
Aloysius Poh Leong
AP
Ting
Grace Kah Mun
GK
Low
Kalpana
K
Gopalakrishnan
M Prakash
MP
Hande
D000328:Adult; D016922:Cellular Senescence / Q000502:physiology; D004249:DNA Damage; D004265:DNA Helicases / Q000235:genetics / Q000378:metabolism; D004260:DNA Repair; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005260:Female; D005347:Fibroblasts / Q000166:cytology* / Q000187:drug effects / Q000502:physiology*; D042822:Genomic Instability; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D008297:Male; D016877:Oxidants / Q000494:pharmacology; D018384:Oxidative Stress; D016615:Telomere / Q000378:metabolism*; D014983:Xeroderma Pigmentosum / Q000235:genetics / Q000378:metabolism
D004268:DNA-Binding Proteins; D016877:Oxidants; C072022:XPBC-ERCC-3 protein; D006861:Hydrogen Peroxide; D004265:DNA Helicases