Protein bound to mitochondrial DNA from tumor and normal tissues of humans and animals.
Stable mitochondrial (mt) DNA-protein complexes have been reported repeatedly in the last decade. We have found that the amount of proteins bound to mt DNA was increased in human HL-60 promyelocytic and chronic myelocytic leukemia cells. Mt of human tumor cells and bovine and rate liver cells were isolated by differential centrifugation. The DNA was purified by SDS-NaCl precipitation of protein, alcohol precipitation, and sucrose density gradient centrifugation. Mt DNA-bound protein obtained by enzymatic digestion of the DNA and purified by gel exclusion and reverse-phase HPLC chomatographies and SDS-polyacrylamide gel electrophoresis, showed a major protein band at 70 kD and a minor band at 35 kD. Hydrolysis of the mtDNA-protein complex in formic acid yielded DNA bases and peptides by HPLC on a C18 reverse-phase column. Tumor cell mtDNA contained 5-10 times more bound protein than mtDNA from normal tissue.
Shapira
N A
NA
Department of Biochemistry, Emory University School of Medicine, Atlanta, Georgia 30322.
Kinkade
J M
JM
Shapira
R
R
D000328:Adult; D000818:Animals; D002417:Cattle; D002499:Centrifugation, Density Gradient; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D004272:DNA, Mitochondrial / Q000378:metabolism*; D004273:DNA, Neoplasm / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006801:Humans; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000378:metabolism*; D015473:Leukemia, Promyelocytic, Acute / Q000378:metabolism*; D008297:Male; D008930:Mitochondria, Liver / Q000378:metabolism; D051381:Rats
D004272:DNA, Mitochondrial; D004273:DNA, Neoplasm