The Repair of DNA Modified by Cytotoxic, Mutagenic, and Carcinogenic Chemicals

Dividing cells from persons with Bloom's syndrome, an autosomal recessive disorder of growth, exhibit increased numbers of chromatid breaks and rearrangements. A highly characteristic feature of the chromosome instability in this syndrome is the tendency for exchanges to occur between chromatids of homologous chromosomes at homologous sites. In the present experiments, a cytogenetic technique by which the sister chromatids of a metaphase chromosome are stained differentially has been used to demonstrate a striking and possibly specific, but hitherto unrecognized, increase in the frequency with which sister chromatids also exchange segments. The cells were grown in bromodeoxyuridine and stained with 33258 Hoechst and Giemsa. Whereas phytohemagglutinin-stimulated lymphocytes from normal controls had a mean of 6.9 sister chromatid exchanges per metaphase (range 1-14), those from persons with Bloom's syndrome had a mean of 89.0 (range 45-162). Normal frequencies of sister chromatid exchanges were found in cells heterozygous for the Bloom's syndrome gene, and also in cells either homozygous or heterozygous for the genes of the Louis-Bar (ataxia telangiectasia) syndrome and Fanconi's anemia, two other rare disorders characterized by chromosome instability. In a differentially stained chromatid interchange configuration discovered during the study, it was possible to determine the new distribution of both sister and non-sister-but-homologous chromatids that had resulted from numerous exchanges. By following shifts in the pattern of staining from chromatid to chromatid, visual evidence was obtained that the quadriradial configurations long recognized as characteristic of Bloom's syndrome represent exchanges between homologous chromosomes, apparently at homologous points. We postulate that the increase in the frequency of exchanges between nonsister-but-homologous chromatids and those between sister chromatids in Bloom's syndrome represents aspects of one and the same disturbance. A study of this phenomenon in relation to the clinical features of Bloom's syndrome may be helpful eventually in understanding the biological significance of chromatid exchange in somatic cells. Show more

We previously described a set of four strains of Salmonella typhimurium designed for detecting the various types of mutagens, and showed their utility in detecting a wide variety of carcinogens as mutagens. The lipopolysaccharide that normally coats these bacteria is a barrier to penetration of mutagens to the cell membrane. The set of tester strains has been improved by adding a mutation (rfa: deep rough) that results in a deficient lipopolysaccharide. The techniques for using these strains for detecting mutagens are presented and the tests are shown to be extremely sensitive and convenient. The specificity of frameshift mutagenesis is clarified. As adjuncts to the test with the four strains, we describe a test that compares mutagenic killing in deep rough strains with and without DNA excision repair, and a test using forward mutagenesis in a deep rough strain lacking excision repair. Show more