DNA replication in isolated HeLa cell nuclei |
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Authors: | O Bernard T P Brent |
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Institution: | McGill University Cancer Research Unit, Montreal, Quebec, Canada |
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Abstract: | DNA replication was investigated in HeLa cell nuclei isolated from different phases of the cell cycle. DNA synthesis occurred only in S-phase nuclei and was dependent on the presence of the four deoxynucleoside triphosphates, Mg++, ATP and S-phase cytoplasm. G1-phase cytoplasm was unable to support such DNA synthesis. A purified preparation of calf thymus DNA polymerase, however, was able to replace S-phase cytoplasm in supporting ATP dependent DNA synthesis, which suggests that the S-phase cytoplasmic factor is a DNA polymerase. G1-phase nuclei could under no conditions be stimulated to initiate DNA replication prematurely. |
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