title: The role of oxygen metabolism for the pathological phenotype of Fanconi anemia.
abstract: The molecular defect of the hereditary disease Fanconi anemia (FA) remains unknown. The two theoretical possibilities are (1) an impaired DNA crosslink-repair system or (2) a disturbed oxygen metabolism either by overproduction of reactive oxygen intermediates (ROI) or by diminished detoxification of ROI. In order to gain further insight into the molecular mechanism of this disease, we have determined the repair capacity of FA cells challenged by crosslinking agents and have analyzed diverse biological systems that are involved in oxygen metabolism. We have tested normal and FA cells for oxygen consumption and for the activity of the antioxidant phospholipid-hydroperoxide-glutathione-peroxidase (PHGPx). FA cells show a reduced oxygen consumption and an increased PHGPx activity. Since spontaneous and induced chromosomal instability is a main cellular feature of FA, we have analyzed the redox state of cells and the effect of cytochrome P-450 (Cyt P-450) inhibitors and inducers on chromosomal breaks and micronuclei production. Our results indicate that Cyt P-450 enzymes, especially Cyt P-450 1A2, play a crucial role in radical metabolism in FA cells. Furthermore, we have determined NF-kappa B activity in untransformed cells and in SV40-transformed cells by gel shift experiments. NF-kappa B is a multiunit transcription factor that is known to be induced by ROI and that activates the expression of various genes involved in cellular responses to stress. NF-kappa B is constitutively induced in SV40-transformed FA cells probably as a consequence of an increased ROI level. Our results suggest that enzymatic defects in oxygen metabolism mediate the FA phenotype via impaired reactivity with ROI. Cyt P-450 1A2 appears to be a good candidate for the defective enzyme, even though no differences have been measured in the activity of this enzyme in FA and control fibroblasts in pilot experiments.
authors.affiliation: Institut für Biochemie, Berlin, Germany.
authors.forename: W
authors.initials: W
authors.lastname: Ruppitsch
authors.forename: C
authors.initials: C
authors.lastname: Meisslitzer
authors.forename: H
authors.initials: H
authors.lastname: Weirich-Schwaiger
authors.forename: H
authors.initials: H
authors.lastname: Klocker
authors.forename: C
authors.initials: C
authors.lastname: Scheidereit
authors.forename: M
authors.initials: M
authors.lastname: Schweiger
authors.forename: M
authors.initials: M
authors.lastname: Hirsch-Kauffmann
mesh_terms: D002460:Cell Line; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D004260:DNA Repair; D018432:Drug Resistance, Multiple / Q000235:genetics; D005199:Fanconi Anemia / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D005347:Fibroblasts / Q000378:metabolism; D005979:Glutathione Peroxidase / Q000378:metabolism; D006801:Humans; D016328:NF-kappa B / Q000378:metabolism; D010084:Oxidation-Reduction; D010100:Oxygen / Q000378:metabolism*; D010101:Oxygen Consumption / Q000235:genetics; D010641:Phenotype; D000080662:Phospholipid Hydroperoxide Glutathione Peroxidase; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: D016328:NF-kappa B; D017382:Reactive Oxygen Species; D003577:Cytochrome P-450 Enzyme System; D000080662:Phospholipid Hydroperoxide Glutathione Peroxidase; D005979:Glutathione Peroxidase; D010100:Oxygen