1.Department of Molecular Biology,Vanderbilt University,Nashville
Abstract:
THE molecular mechanism for initiating DNA replication can be studied using a subcellular system. Rao and Johnson1 found that HeLa cells in the pre-DNA-synthetic (G-1) period of the cell cycle initiate DNA synthesis after fusion with cells that are in the DNA synthetic (S) period. A previous subcellular system of DNA replication from HeLa cells2–4 consisted of intact nuclei, supplemented with the four deoxy-nucleoside triphosphates, salt, ATP and a cytosol factor. The nuclei in this system appeared to be permeable to proteins and DNA synthesis was very similar to that within intact cells. We report here the initiation of DNA synthesis in nuclei isolated from HeLa cells. Our results suggest that, with the synchronization method used, a small percentage of dormant G-1 nuclei can be stimulated by S-phase cytoplasm; this would be the case if the cells were receptive to stimulation for only 30–60 min during the cell cycle. illustration