Evidence for the existence of a stable association between nascent DNA and the nuclear membrane of HeLa cells.
Nascent DNA-nuclear membrane complexes isolated from HeLa cells and solubilized in a sodium dodecyl sulfate-urea solution were examined by gel electrophoresis, column chromatography, isopycnic centrifugation, and by extraction with chloroform/methanol. Radioactivity attributable to [3H]DNA co-migrated with three protein peaks during electrophoresis. This radioactivity was eliminated by prior treatment with DNAase. In addition, all of the radioactivity attributable to nascent DNA eluted with a specific protein on Sepharose 4B columns. This DNA - protein complex banded at a density of 1.58 gm/cm3 in sucrose-CsCl gradients. Treatment with DNAase, phospholipase A and C, and dilute alkali disrupted the complex. Moreover, 93% of the radioactivity attributable to protein and 70% of that attributable to DNA could be extracted from the complex with a chloroform/methanol solution. The results suggest that nascent DNA may be in a stable association with a proteolipid moiety of the nuclear membrane.
D001665:Binding Sites; D002455:Cell Division; D002467:Cell Nucleus / Q000378:metabolism*; D002499:Centrifugation, Density Gradient; D004273:DNA, Neoplasm / Q000096:biosynthesis / Q000378:metabolism*; D006367:HeLa Cells / Q000378:metabolism*; D008566:Membranes / Q000378:metabolism; D014176:Protein Biosynthesis; D011955:Receptors, Drug
D004273:DNA, Neoplasm; D011955:Receptors, Drug