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Localization of chromosome breakpoints induced by DNase I in Chinese hamster ovary (CHO) cells
1División de Citogenética Humana y Microscopía Cuantitativa, Instituto de Investigaciones Biológicas Clemente Estable, Avenida Italia 3318, C.P. 11600, Montevideo, Uruguay. folle@iibce.edu.uy
Abstract:
DNase I was electroporated into S-phase CHO cells and induced chromosome breakpoints were localized in G-banded metaphases. More than 75% of breakpoints mapped to Giemsa-light bands, 18% to Giemsa-dark bands and about 7% to band junctions. Chromosome breakpoint clusters produced by DNase I colocalized with chromosome breakpoints induced by the restriction endonucleases AluI and BamHI in the G1- and S-phases of the cell cycle in CHO cells. Digestion of metaphase spreads with AluI, BamHI and DNase I produced G-bands, indicating that G-light bands are more sensitive to endonuclease action. The possible role of nuclease-sensitive sites in active chromatin as selective targets for the induction of chromosome breakpoints by these endonucleases is discussed.