title: DNA repair fine structure in Werner's syndrome cell lines.
abstract: Werner's syndrome (WS) is a human segmental progerioid disorder with an autosomal recessive pattern of inheritance. Patients with WS exhibit a number of symptoms resembling a premature aging phenotype. We have examined the fine structure of the DNA repair of UV-induced cyclobutane pyrimidine dimers in Epstein-Barr virus (EBV)-transformed WS lymphoblastoid cell lines and in a primary WS fibroblast cell line. The repair was measured at the level of the gene and also in the general genome. Gene-specific and strand-specific DNA repair was measured in the actively transcribed genes dihydrofolate reductase (DHFR), c-myc, and p53, and in the transcriptionally inactive regions, delta globin and the X-linked 754 domain. Both gene-specific repair and strand-specific repair were deficient in the transformed WS lymphoblastoid cell lines compared to normal controls. In normal cells, repair in the transcribed strand was 25 (4 h), 43 (8 h), and 72% (24 h); in the WS cells on average, repair in the transcribed strand was 18 (4 h), 27 (8 h), and 44% (24 h). However, in the primary WS fibroblast cell line, we found a pattern of preferential gene repair which was similar to that in normal human cells. In contrast to cells from patients with the gene-specific repair deficient disease Cockayne's syndrome, which show greatly delayed RNA synthesis recovery after UV irradiation, the WS cells had normal recovery of RNA synthesis. The DNA repair results differ for the different cell types, and our findings thus do not establish a general DNA repair phenotype for WS cells. The fibroblasts had proficient repair, but in the WS lymphoblasts we find a deficiency in DNA repair which could contribute to the reported hypermutability in these cells. The lymphoblasts are, however, transformed cells, and it raises the concern that biological findings in transformed cells may not reflect the situation in primary cells.
authors.affiliation: Laboratory of Molecular Genetics, National Institute on Aging, National Institutes of Health, Baltimore, MD 21224, USA.
authors.forename: D K
authors.initials: DK
authors.lastname: Webb
authors.forename: M K
authors.initials: MK
authors.lastname: Evans
authors.forename: V A
authors.initials: VA
authors.lastname: Bohr
mesh_terms: D015139:Blotting, Southern; D002461:Cell Line, Transformed / Q000166:cytology / Q000201:enzymology; D004260:DNA Repair / Q000502:physiology* / Q000528:radiation effects; D004277:DNA, Single-Stranded / Q000502:physiology* / Q000528:radiation effects; D005347:Fibroblasts / Q000166:cytology / Q000201:enzymology; D004854:Herpesvirus 4, Human / Q000235:genetics; D006801:Humans; D008214:Lymphocytes / Q000166:cytology / Q000201:enzymology; D012313:RNA / Q000096:biosynthesis; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics; D014158:Transcription, Genetic / Q000502:physiology; D014466:Ultraviolet Rays; D014898:Werner Syndrome / Q000201:enzymology / Q000235:genetics*
chemical_list: D004277:DNA, Single-Stranded; D012313:RNA; D013762:Tetrahydrofolate Dehydrogenase