CpG Islands and Double-Minute Chromosomes

Abstract

Double-minute chromosomes (DMs) amplify oncogenes in human tumors. The organization of genomic DNA in four independently isolated DMs amplifying the DHFR (dihydrofolate reductase) gene has been compared by mapping locations of CpG islands. When cleaved with methylation-sensitive rare-cutting restriction endonucleases, three hypomethylated GC-rich DNA sequences were frequently found in specific regions in these DMs. One such zone was in the CpG island containing the divergently transcribed promoter separating the DHFR and the Rep-3 genes. The other two sites were approximately 500 kb upstream and 300 kb downstream of the DHFR gene. An approximately 800-kb amplified core genomic region containing the DHFR gene using DM-specific probes has been identified in this study. All the DMs consisted of the core amplified region combined with additional DNA fragments. These additional fragments are different for each DM. Therefore, while the DNAs in each of the DMs are different, they have common hypomethylated regions in similar locations. These results suggest a role for the location of hypomethylated GC-rich sites such as the CpG islands in genesis of DMs.