Patterns of BrdU incorporation in homogeneously staining regions and double minutes

The replication chronology of two structural chromosome abnormalities linked to the amplification phenomenon of DNA sequences was investigated. Three cell lines containing homogeneously staining region (HSR) chromosomes (IMR-32, MK42, and COLO-320) and one line with double minutes (DM) (SW-613) were examined. Using a bromodeoxyuridine-Hoechst 33258-Giemsa method, the HSR in the three cell lines were shown to be composed of subunits that replicated their DNA throughout all portions of the S-phase of the cell cycle. The double minute chromosomes were observed to replicate randomly throughout the entire S-phase, with no pattern evident. These results are consistent with the suggestion that DNA from HSR and DM are structurally and functionally related. Moreover, this observation that these amplified regions replicate their DNA throughout the entire S-phase favors the idea that, during amplification processes, both early and late replicating sequences are included. The apparent discordance between staining characteristics and replication behavior exhibited by some HSR and DM are also discussed.