Study of HL-60 cells DNA synthetic machinery: Isolation, characterization and its biochemical status at differential cellular growth stages

The mammalian cell DNA synthetic machinery has been isolated from human cervical carcinoma HeLa and murine FM3A cells, and characterized as a multiprotein replication complex (MRC). This complex is fully capable of supporting SV40 in vitro DNA replication in the presence of viral large T antigen. To further study the regulation of the activity of the MRC during cell differentiation, a human leukemia HL-60 cell model was used. HL-60 cells are able to undergo macrophage-like differentiation after exposure to a phorbol ester. The differentiation is accompanied by the cessation of DNA synthesis. In the attempt to isolate and characterize the HL-60 cell DNA synthetic machinery, we found that several essential DNA replication proteins such as DNA polymerases...