Evidence for the existence of a stable association between nascent DNA and the nuclear membrane of HeLa cells.

Biochimica Et Biophysica Acta
D M Dye, A P Toliver

Abstract

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.

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Citations

Oct 1, 1995·The International Journal of Biochemistry & Cell Biology·T Neitcheva, D Peeva
Apr 1, 1979·International Journal of Radiation Biology and Related Studies in Physics, Chemistry, and Medicine·A W KoningsW B Trieling
Apr 1, 1977·International Journal of Radiation Biology and Related Studies in Physics, Chemistry, and Medicine·A W Konings, W B Trieling
Apr 10, 1978·Biochimica Et Biophysica Acta·J R Harris

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