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Two- and Three-Dimensional Live Cell Imaging of DNA Damage Response Proteins
Published on: September 28, 2012
Active dissociation of the fluorescent dye Hoechst 33342 from DNA in a living cell: who could do it?
S N Naryzhny1, V V Levina, E Y Varfolomeeva
1Petersburg Nuclear Physics, Institute of Russian Academy of Sciences, Gatchina, Leningrad district. naryzhyny@omrb.pnpi.spb.ru
Abstract:
It is assumed that DNA in mammalian cells is a dynamic conformationally unstable system. This instability provides the cell with a mechanism for dissociating a large number of substances that bind tightly but not covalently to DNA. Among these is the fluorescent dye Hoechst 33342, which binds to DNA in the minor groove. We have selected cell lines with a high capability for active dissociation of Hoechst 33342. Comparative protein analysis of these lines by means of two-dimensional (2-D) electrophoresis was performed. Cell and nuclear proteins were analyzed from these and normal strains. A few proteins with significantly changed quantities have been found. The preliminary search of the 2-D database allowed us to identity some known and unknown cellular proteins that could participate in active dissociation of the dye from DNA.
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