We established the size distribution of extrachromosomal covalently closed circular DNA molecules from embryos of various Drosophila melanogaster strains and from Kc0% tissue culture cells. In embryos, more than 80% of the circular DNA molecules are smaller than 2.5 kb and all the distributions show a peak of molecules of between 200 and 400 bp. The Kc0% cell distribution differs mainly from that of embryos in that 48% of the molecules have a size between 4 and 8 kb. Correlating with this, circular molecules homologous to copia, 412 and 297 were detected only in Kc0% cells. The three tandemly repeated families containing the 5S genes, the histone genes and the 240 bp repeat of the ribosomal DNA intergenic spacer, which had previously been identified in circular DNAs from embryos, were also found in cultured cells. A fourth tandemly repeated family corresponding to the 1.688 g/cm3 satellite DNA was detected, both in embryos and Kc0% cells. It consists of circular multimeric molecules containing multiple copies of the 359 bp repeated unit. No circular DNA molecules homologous to the actin genes, the type I ribosomal DNA insertion, or the F and I transposable elements were found in embryos or Kc0% cells. Thus it appears that the extrachromosomal circular DNA molecules from embryos and from tissue culture cells differ mainly in the presence of circular copies of the copia-like transposable elements.

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Used at a physiological concentration, the steroid hormone 20-hydroxyecdysone (20-OHE) induces, in Kc cultured Drosophila melanogaster cells, important and specific changes. Modifications occur at morphological and enzymatical levels. Variations in specific protein synthesis are observed. At the molecular level, 20-OHE particularly induces a decrease in expression of the mobile dispersed genetic element 412. This repeated element which belongs to the "copia-like" family is more widely represented in Kc cells (80 fold) compared to fly cells (25 fold). 412 transcripts are heterogeneous in size, essentially polyadenylated and restricted to the nucleus. A minimal concentration of 10(-8) M and a time treatment of 16 hours are necessary to obtain a strong decrease in 412 expression. The decrease is at least an effect on these sequences at the transcriptional level. Structural similarities between the 412 element and the proviral forms of vertebrate retroviruses are strengthened by the characterization of extrachromosomal circular DNA forms revealed by the 412 probe. Quantifying experiments have shown that the steady state level of such forms is not affected by the steroid treatment.

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