Co-localization of mitochondrial and double minute DNA in the nuclei of HL-60 cells but not normal cells.
In an attempt to isolate genes located on double minute (Dmin) DNA in HL-60 cells, we prepared DNA probe from purified micronuclei. Micronucleation was induced in HL-60 cells by treatment with hydroxyurea. Screening of a cDNA library unexpectedly produced a number of clones containing mitochondrial DNA (mtDNA) sequences. Here, we show that amplified mtDNA sequences were localized in nuclei and micronuclei of HL-60 and COLO 320DM cells, but not in nuclei of WI-38 normal human fibroblasts or peripheral blood T-cells. To unequivocally demonstrate the presence of mtDNA inside of nuclei and micronuclei, we obtained tomographic fluorescence in situ hybridization (FISH) images of mtDNA by confocal microscopy of consecutive sections of paraformaldehyde (PFA)-fixed material. We also located mtDNA in nuclear buds and purified micronuclei. Dmin DNA and mtDNA were always located at similar sites. The mechanisms of nuclear retention of mtDNA and Dmin DNA and the resulting influence on tumorigenesis are discussed.
Hirano
T
T
Life Science Group, Faculty of Integrated Arts and Sciences, Hiroshima University, Kagamiyama 1-7-1, Higashihiroshima 739-8521, Japan.
Shiraishi
K
K
Adachi
K
K
Miura
S
S
Watanabe
H
H
Utiyama
H
H
D002453:Cell Cycle / Q000187:drug effects; D002467:Cell Nucleus / Q000737:chemistry*; D002471:Cell Transformation, Neoplastic / Q000235:genetics; D003110:Colonic Neoplasms / Q000473:pathology; D004249:DNA Damage; D004272:DNA, Mitochondrial / Q000032:analysis*; D004273:DNA, Neoplasm / Q000032:analysis*; D005347:Fibroblasts / Q000737:chemistry / Q000648:ultrastructure; D005784:Gene Amplification; D018922:HL-60 Cells / Q000737:chemistry / Q000187:drug effects / Q000648:ultrastructure*; D006801:Humans; D006918:Hydroxyurea / Q000494:pharmacology; D017404:In Situ Hybridization, Fluorescence; D048629:Micronuclei, Chromosome-Defective / Q000737:chemistry / Q000187:drug effects / Q000648:ultrastructure*; D018613:Microscopy, Confocal; D013601:T-Lymphocytes / Q000737:chemistry / Q000648:ultrastructure; D014407:Tumor Cells, Cultured / Q000737:chemistry / Q000648:ultrastructure
D004272:DNA, Mitochondrial; D004273:DNA, Neoplasm; D006918:Hydroxyurea