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. 1997 Feb 24;231(3):856-60.
doi: 10.1006/bbrc.1997.6203.

In organello footprinting analysis of rat mitochondrial DNA: protein interaction upstream of the Ori-L

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In organello footprinting analysis of rat mitochondrial DNA: protein interaction upstream of the Ori-L

G Cingolani et al. Biochem Biophys Res Commun. .

Abstract

An in organello footprinting approach has been used to probe a protein-DNA interaction of a nuclear coded 25 kDa protein, previously isolated in our laboratory, that binds "in vitro" a region within the ND2 gene, located upstream of the Ori-L. Footprinting studies with the purine-modifying reagent dimethyl sulfate and the pirimidine-modifying reagent potassium permanganate were carried out in isolated mitochondria from rat liver. Dimethyl sulfate footprinting has allowed the detection of a protein-DNA interaction within the curved ND2 region with contact sites located in both the strands. Potassium permanganate footprinting allowed detection of an adjacent permanganate-reactive region. We hypothesize that the permanganate-reactive region is a single stranded DNA due to a profound helix distortion induced by a 25 kDa protein binding to the nearest region.

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