In organello footprinting analysis of rat mitochondrial DNA: protein interaction upstream of the Ori-L.
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.
Cingolani
G
G
Department of Biochemistry and Molecular Biology, University of Bari, Italy.
Capaccio
L
L
D'Elia
D
D
Gadaleta
G
G
D000818:Animals; D001483:Base Sequence; D018983:DNA Footprinting; D004261:DNA Replication; D004272:DNA, Mitochondrial / Q000378:metabolism* / Q000648:ultrastructure; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism; D003852:Deoxyribonucleoproteins / Q000737:chemistry; D008930:Mitochondria, Liver / Q000737:chemistry / Q000648:ultrastructure*; D008970:Molecular Weight; D009690:Nucleic Acid Conformation; D011196:Potassium Permanganate / Q000737:chemistry; D011485:Protein Binding; D051381:Rats; D013463:Sulfuric Acid Esters / Q000737:chemistry
D004272:DNA, Mitochondrial; D004268:DNA-Binding Proteins; D003852:Deoxyribonucleoproteins; D013463:Sulfuric Acid Esters; D011196:Potassium Permanganate; C007482:dimethyl sulfate