In Drosophila, the as yet uncloned heterochromatic locus flamenco (flam) controls mobilization of the endogenous retrovirus gypsy through the repeat-associated small interfering (rasi) RNA silencing pathway. Restrictive alleles (flamR) downregulate accumulation of gypsy transcripts in the somatic follicular epithelium of the ovary. In contrast, permissive alleles (flamP) are unable to repress gypsy. DIP1, the closest transcription unit to a flam-insertional mutation, was considered as a good candidate to be a gypsy regulator, since it encodes a dsRNA-binding protein. To further characterize the locus we analyzed P-induced flam mutants and generated new mutations by transposon mobilization. We show that flam is required somatically for morphogenesis of the follicular epithelium, the tissue where gypsy is repressed. This developmental activity is necessary to control gypsy and another retroelement, ZAM. We also show that flam is not DIP1, as none of the new permissive mutants affect the DIP1 coding sequence. In addition, two deletions removing DIP1 coding sequences do not affect any of the flamenco functions. Our results suggest that flamenco extends proximally to DIP1, spanning >130 kb of transposon-rich heterochromatin. We propose a model explaining the multiple functions of this large heterochromatic locus.

We have used a combination of a sensitive immunocytochemical stain for intracellular albumin, and Hoechst 33258 dye for identification of parental nuclei to investigate the time-course of extinction, reexpression, and activation of albumin production in fusion products of 1s (hyperdiploid) or 2s (hype rtetradiploid) rat hepatoma cells with mouse fibroblasts (L cells or embryonic cells) . In all combinations, the initial event is extinction of albumin production . Extinction occurs immediately after fusion when the mouse fibroblast is a normal embryonic (senescent?) cell . In the case of an L cell, rat albumin is synthesized and secreted during the first 12 h after fusion ; no production of mouse albumin occurs . Thereafter, albumin production ceases . 8-12 d after fusion, young hybrid colonies are found to resume the synthesis of rat albumin (reexpression), and several days later the production of mouse albumin begins (activation) . The patterns of reexpression and activation indicate (a) that chromosome loss is not necessary for either event to occur and (b) that the cells active in the synthesis of mouse albumin are a subpopulation of those cells already engaged in the production of rat albumin. We conclude that (a) extinction is mediated by diffusible factor(s) from the L-cell parent that act in the hepatoma nucleus to prevent the formation of new albumin messenger RNA; (b) reexpression and activation are gene dosage-dependent but extinction is not; and (c) previously active genes are more rapidly expressed than previously silent ones .

The ovo locus is required for the maintenance of the female germ line in Drosophila melanogaster. In the absence of an ovo+ gene, males are completely normal but females have no germ‐line stem cells. Three dominant mutations at the ovo locus, called ovoD, were observed to revert towards recessive alleles at high frequency when ovoD males were crossed to females of the strain y v f mal. We have found that this strain contains an inordinately high number of gypsy transposable elements, and crossing it with the ovoD strains results in the mobilization of both gypsy and copia, with high‐frequency insertions into the ovo locus: of 16 revertants examined 12 have gypsy and four have copia inserted at 4E, the ovo cytological site. Using gypsy DNA as a tag we have cloned 32 kb of wild‐type DNA sequences surrounding a gypsy insertion and characterized molecular rearrangements in several independent revertants: in 10 of them gypsy appears to be inserted into the same site. The orientation of gypsy is strictly correlated with whether the neighbouring lozenge‐like mutation appears in the revertants. A distal limit of the ovo locus was molecularly determined from the breakpoint of a deletion affecting closely flanking regions.

As defined by dominant and recessive ovo mutations, the ovo gene is required for development of the Drosophila female germ line, and does not exert any function in males or in somatic tissues. However, reversion of dominant ovo mutations can result in new phenotypes that are not related to the female germ line: the svb and lzl mutations affect cuticle and eye development, respectively. We have identified a 7.2 kb genomic fragment that rescues ovo mutations in transgenic Drosophila and thus contains all sequences necessary for ovo+ function. This fragment has been sequenced almost in its entirety, defining the ovo locus at the molecular level. Multiple copies of the same fragment also rescue the lzl mutation. They do not rescue svb mutations, in agreement with genetic evidence that the svb function requires additional, more distal sequences. Nevertheless, a number of transposable element insertions that induce a svb phenotype interrupt the coding sequence of ovo. Taken together, the genetic and molecular data indicate the existence of a complex locus, where the ovo and svb functions depend on overlapping coding sequences but distinct regulatory elements. The data also suggest a model for the lzl phenotype. Expression of ovo at the RNA level is detectable at stage 8 of oogenesis in nurse cells and persists through the rest of oogenesis and in early embryogenesis. The ovo transcript encodes a protein of at least 1209 amino acids with four zinc fingers, suggesting that ovo might be a transcription factor required for female germ line maintenance and gametogenesis.