Journal
CYTOMETRY PART A
Volume 73A, Issue 6, Pages 555-562Publisher
WILEY-LISS
DOI: 10.1002/cyto.a.20573
Keywords
chromatin structure; histone exchange; DNA intercalator; minor groove binder; DRAQ5; Syto17; fluorescence confocal microscopy
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DNA-binding dyes are useful in contrasting cell nuclei and chromatin for live cell fluorescence rnicroscopy; however, they may interfere with nuclear and DNA structure and function. We investigated the influence exerted by two DNA dyes on chromatin and nuclear structure, as well as histone-DNA interactions in live HeLa cells. A membrane-permeant fluorescent DNA intercalator DRAQ5 (anthracycline derivative), at a concentration of 1 mu M, caused microscopically detectable changes of nuclear architecture. Following DRAQ5 intercalation into DNA, chromatin aggregated into distinct areas and foci. The loss of 3D chromatin distribution was exerted via interference with a dynamic exchange of a linker histone (H1), which is a known chromatin stabilizing factor. At higher concentrations (3 and 7.5 mu M), DPAQ5 interfered with binding of H2B core histones to DNA. Similar effects resulted from intercalation of chemotherapeutic drugs, adriamycin and daunomycin, but were not observed after binding to DNA of a minor groove binder, Syto17. (c) 2008 International Society for Advancement of Cytometry.
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