The dynamic distribution and quantification of DNA cruciforms in eukaryotic nuclei.
Cruciforms have been suggested as potential recognition structures at or near origins of DNA replication in eukaryotic cells. Monoclonal antibodies specific for cruciforms have been produced. The antibody binds to structural determinants at the base of the cruciform stem, the "elbow." Labeling of nuclei with anti-cruciform antibodies produces a nonuniform pattern of fluorescence in cells arrested at the G1/S boundary. This pattern of fluorescence changes when these cells are released from synchrony. Using fluorescence flow cytometry to quantify the number of DNA cruciform structures in cells throughout the cell cycle, we observed two major populations of nuclei with different numbers of cruciforms; the modal number of cruciforms in these populations was 0.6 x 10(5) and 3 x 10(5) cruciforms per nucleus. Synchronized cells (doubly arrested by serum starvation and aphidicolin) displayed a biphasic distribution of the number of cruciforms over the first 6 h after release from synchrony with maxima at 0 and 4 h after release.
McGill Cancer Center, McGill University, Montreal, Quebec, Canada.
G K
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Ward
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McKenzie
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Zannis-Hadjopoulos
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Price
D000818:Animals; D002453:Cell Cycle; D002460:Cell Line; D002467:Cell Nucleus / Q000032:analysis*; D002469:Cell Separation; D004247:DNA / Q000032:analysis* / Q000276:immunology; D004261:DNA Replication; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D006801:Humans; D007399:Interphase; D009690:Nucleic Acid Conformation
D004247:DNA