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Radiation-Induced Association of β-Glucuronidase with Purified Nuclei from Irradiated MOLT-4 and HeLa Cells
D. E. McClain, J. F. Kalinich, J. K. Poplack and S. L. Snyder
Vol. 117, No. 2 (Feb., 1989), pp. 221-233
Published by: Radiation Research Society
Stable URL: http://www.jstor.org/stable/3577323
Page Count: 13
You can always find the topics here!Topics: Cell lines, HeLa cells, Cell nucleus, Enzymes, Lysosomes, Incubation, Radiation dosage, Nuclear membrane, Radiation damage, Lymphocytes
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Beta-glucuronidase, a lysosomal marker enzyme, associates with purified nuclei from HeLa and MOLT-4 cell lines in a radiation dose-dependent manner, up to 300 cGy in MOLT-4 cells, and 1000 cGy in HeLa cells. In MOLT-4 cells (200-cGy exposure), there is a significant increase in β-glucuronidase activity detected in the nuclear fraction 24 h postirradiation with a maximum association occurring at 72 h. In HeLa cells (1000-cGy exposure), a significant association is first detected 24 h postirradiation with a maximum association at 48 h. The association is not the result of nonspecific contamination occurring during nuclei purification since nuclei from irradiated cells show no greater levels of plasma membrane marker and mitochondrial marker than controls. The nature of the association remains unclear, but activity is not removed by detergents used in the nuclei isolation procedure, and incubation of the nuclei with EDTA reverses the association only modestly. Exposure of nuclei from irradiated cells to anisotonic buffers also results in only a small decrease in β-glucuronidase activity associated with the nuclei. These observations suggest that lysosomal hydrolases become intimately associated with the nuclei of irradiated cells.
Radiation Research © 1989 Radiation Research Society