Localization of specific sequences and DNA single-strand breaks in individual UV-A-irradiated human lymphocytes by COMET FISH

Claudia Bock; Alexander Rapp; Heike Dittmar; Shamci Monajembashi; Karl-Otto Greulich

doi:10.1117/12.336836

19 January 1999 Localization of specific sequences and DNA single-strand breaks in individual UV-A-irradiated human lymphocytes by COMET FISH

Claudia Bock, Alexander Rapp, Heike Dittmar, Shamci Monajembashi, Karl-Otto Greulich

Author Affiliations +

Proceedings Volume 3568, Optical Biopsies and Microscopic Techniques III; (1999) https://doi.org/10.1117/12.336836
Event: BiOS Europe '98, 1998, Stockholm, Sweden

Abstract

The COMET assay, a single cell electrophoresis technique which allows to separate electrophoretically fractionated DNA according to size has been combined with fluorescence in situ hybridization (FISH) which allows to localize specific genes or gene regions. This combination (COMET FISH) allows the detection of DNA single strand breaks in specific regions of the genome of human lymphocytes at the single cell level. Various types of DNA probes, e.g. centromere-, (alpha) - satellite-, telomere-, whole chromosome-, single copy- and region specific DNA probes have been used to investigate whether the UV-A induced DNA single strand breaks are distributed randomly all over the human genome or induced at specific sites ('hot spots'). In the investigated human peripheral blood lymphocytes all but one centromere reveal low sensitivity for UV-A irradiation (500 kJ/m²), while telomeres are randomly distributed over COMET heads and tails. The human chromosome 1 is fractionated by irradiation, but remains in the COMET head, indicating an only moderate degree of fractionation. Among three tested single copy probes, c- myc, p53 and p58, the p53 gene located on chromosome 17p13.1 and the p58 gene (1p36) appear to be located in UV-A stable regions of the human genome in 95% of 65 investigated lymphocytes. In contrast, the c-myc proto-oncogene (8q24) is found in the COMET tail in 90% of the 27 investigated lymphocytes and thus appears to be more sensitive to UV-A irradiation.

Citation Download Citation

Claudia Bock, Alexander Rapp, Heike Dittmar, Shamci Monajembashi, and Karl-Otto Greulich "Localization of specific sequences and DNA single-strand breaks in individual UV-A-irradiated human lymphocytes by COMET FISH", Proc. SPIE 3568, Optical Biopsies and Microscopic Techniques III, (19 January 1999); https://doi.org/10.1117/12.336836

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