RESUMO
The well-known sex-determining cascade of Drosophila melanogaster serves as a paradigm for the pathway to sexual development in insects. But the primary sex-determining signal and the subsequent step, Sex-lethal (Sxl), have been shown not to be functionally conserved in non-Drosophila flies. We isolated doublesex (dsx), which is a downstream step in the cascade, from the phorid fly Megaselia scalaris, which is a distant relative of D. melanogaster. Conserved properties, e.g., sex-specific splicing, structure of the female-specific 3' splice site, a splicing enhancer region with binding motifs for the TRA2/RBP1/TRA complex that activates female-specific splicing in Drosophila, and conserved domains for DNA-binding and oligomerization in the putative DSX protein, indicate functional conservation of dsx in M. scalaris. Hence, the dsx step of the sex-determining pathway appears to be conserved among flies and probably in an even wider group of insects, as the analysis of a published cDNA from the silkmoth indicates.
Assuntos
Sequência Conservada/genética , Proteínas de Ligação a DNA/genética , Dípteros/genética , Proteínas de Drosophila , Proteínas de Insetos/genética , Splicing de RNA/genética , Processos de Determinação Sexual , Sequência de Aminoácidos , Animais , Sequência de Bases , DNA Complementar/análise , Proteínas de Ligação a DNA/classificação , Evolução Molecular , Éxons , Dosagem de Genes , Proteínas de Insetos/classificação , Íntrons , Dados de Sequência Molecular , Filogenia , RNA/análise , Sequências Reguladoras de Ácido Nucleico , Homologia de Sequência de Aminoácidos , Homologia de Sequência do Ácido NucleicoRESUMO
Sex-lethal (Sxl) and doublesex (dsx) are known to represent parts of the sex-determining cascade in Drosophila melanogaster. We generated cDNA probes of the homologous genes from Megaselia scalaris, a fly species with an epistatic maleness factor as the primary sex determining signal. In Northern blot hybridization of poly(A)+ RNA, the M. scalaris dsx probe detected two bands, one of which had a sex-specific size difference, while the Sxl probe bound to RNAs of equal size in females and males. RT-PCR showed Sxl to be transcribed in gonads of adult females and males but not in somatic tissues. Thus, while dsx appears to have a similar function in M. scalaris and D. melanogaster, Sxl does not. The results suggest that the sex-determining pathway of M. scalaris joins that of D. melanogaster between the Sxl and dsx steps.
RESUMO
Sex-lethal (Sxl) is Drosophila melanogaster's key regulating gene in the sex-determining cascade. Its homologue in Megaselia scalaris, the chromosome 3 gene Megsxl, codes for a protein with an overall similarity of 77% with the corresponding D. melanogaster sequence. Expression in M. scalaris, however, is very unlike that in D. melanogaster. Megsxl transcripts with a long ORF occur in both sexes. Differential splicing is conserved but not sex-specific. There are several splice variants, among them one is common to gonads and somatic tissues of all developmental stages investigated, one is specific for ovaries and embryos, and a third one is not found in ovaries. In the ovary, Megsxl is heavily transcribed in nurse cells and transported into eggs. These results suggest a non-sex-determining function during early embryogenesis; the presence of Megsxl RNA in testes and somatic tissues calls for other (or more) functions.