Amplified DNA sequences in Y1 mouse adrenal tumor cells: association with double minutes and localization to a homogeneously staining chromosomal region.

DNA associated with double minutes (dm) of the Y1-DM mouse adrenocortical tumor cell line has been cloned in Charon 4A and a preliminary characterization has been made of a recombinant clone, lambda Y1dm-1, isolated from this dm DNA library [George, D. L. & Powers, V. E. (1981) Cell 24, 117--123]. Cloned sequences in lambda Y1dm-1 are amplified in the genome of the Y1-DM cells. They are also amplified in the genome of a related Y1 subline (Y1-HSR), which has a homogeneously staining chromosomal region (HSR). Here we report that the amplified sequences complementary to lambda Y1dm-1 are localized in the HSR, as determined by in situ hybridization. In addition, we found that a population of Y1-DM cells originally containing only dm later consisted of two cell types. Some cells retained dm; others had lost dm but gained a HSR-bearing chromosome morphologically distinct from that in the Y1-HSR cell line. Subclones isolated from this mixed culture have either dm or a HSR, but not both. Southern blotting studies revealed that genomic DNA samples from subclones with a HSR, like subclones with dm, still possess amplified copies of DNA homologous to our recombinant probe. These experiments provide direct evidence that the dm and HSRs in these Y1 cells are structurally related and further support the hypothesis that these chromosomal anomalies result from a process of gene amplification.