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In Drosophila Kc cells, the 60C beta3 tubulin transcription unit, whose expression is induced by 20-hydroxyecdysone (20-OHE), has the same structure as in Drosophila. This gene is characterized by an unusual 5' intron of regulating importance, by an alternatively spliced second intron and by a long 3' transcribed but untranslated region. This gene codes for two beta3 tubulin isoforms with one amino acid difference. We have established that beta3 tubulin gene expression is transcriptionally regulated by the steroid hormone in a time and hormonal concentration-dependent fashion, without requirement of protein synthesis. This implies that this transcriptional induction is a primary event and that this gene is probably a direct target for the 20-OHE receptor.

Cultured Drosophila melanogaster cells are responsive to the steroid hormone 20-hydroxyecdysone (ecdysterone). The transcripts of two copia-like transposable elements, 412 and 1731 were examined in cells treated for different lengths of time by the hormone. Using dot-blot and Northern-blot analysis, we have shown that the transcripts of these two retrotransposons were rapidly decreased by ecdysterone treatment.

Cultured Kc cells of Drosophila melanogaster are sensitive to the insect moulting hormone, 20-hydroxy-ecdysone (20-OH-E). Morphological changes of Kc-treated cells were observed and electron microscopic analysis of pseudopodia shows a large increase in the number of microtubules, all arranged in the same orientation. The 60 C beta tubulin gene which is expressed only in 20-OH-E-treated cells encodes a 2.6-kb mRNA which is essentially cytoplasmic and polyadenylated. The corresponding premessenger is 7 kb in length and is absent in untreated cells. Two peaks of expression of the 60 C beta tubulin gene are observed during Drosophila development: at midembryogenesis (stage 8-13 h) and at the late third instar larvae-early pupae stage. By use of the Ecdysone 1 mutant, 60 C beta tubulin gene expression was demonstrated to be regulated in part by 20-OH-E during Drosophila development. Through these two complementary biological models of study, the mode and role of beta tubulin gene regulation are discussed.

Bacterial resistance to mercury has been studied in two different strains from animal origin, Salmonella typhimurium 9205 and Escherichia coli 467. These two strains are resistant to mercuric chloride but sensitive to phenylmercury, and thus belong to the group of bacteria that possess a "narrow" spectrum resistance. The presence of plasmids within the cells has been demonstrated through conjugation experiments and direct detection of extrachromosomal DNA in transconjugants. These plasmids, termed p9205-1 and p467-30, differ from each other by both their size (100 and 50 Kbp, respectively) and structure, as shown by the restriction patterns arising from digestion by nucleases BamHI, HindIII, PstI and EcoRI. Each plasmid has been treated with enzyme SalI to yield DNA fragments that have been cloned into pBR322. Two recombinant plasmids, p9205-1/Sal and p467-30/Sal, have thus been constructed, each of them harboring a 2 Kbp fragment that appears to contain the merA gene coding for mercuric reductase. From the analysis of the restriction maps of these recombinant plasmids as well as the functional behaviour of the bacteria that they are able to transform, it can be concluded that they are identical.

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.