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Background  In the model system  Drosophila melanogaster, doublesex  ( dsx ) is the double-switch gene at the bottom of the somatic sex determination cascade that determines the differentiation of sexually dimorphic traits. Homologues of  dsx  are functionally conserved in various dipteran species, including the malaria vector  Anopheles gambiae . They show a striking conservation of sex-specific regulation, based on alternative splicing, and of the encoded sex-specific proteins, which are transcriptional regulators of downstream terminal genes that influence sexual differentiation of cells, tissues and organs.    Results  In this work, we report on the molecular characterization of the  dsx  homologue in the dengue and yellow fever vector  Aedes aegypti  ( Aeadsx ).  Aeadsx  produces sex-specific transcripts by alternative splicing, which encode isoforms with a high degree of identity to  Anopheles gambiae  and  Drosophila melanogaster  homologues. Interestingly,  Aeadsx  produces an additional novel female-specific splicing variant. Genomic comparative analyses between the  Aedes  and  Anopheles dsx  genes revealed a partial conservation of the exon organization and extensive divergence in the intron lengths. An expression analysis showed that  Aeadsx  transcripts were present from early stages of development and that sex-specific regulation starts at least from late larval stages. The analysis of the female-specific untranslated region (UTR) led to the identification of putative regulatory  cis -elements potentially involved in the sex-specific splicing regulation. The  Aedes dsx  sex-specific splicing regulation seems to be more complex with the respect of other dipteran species, suggesting slightly novel evolutionary trajectories for its regulation and hence for the recruitment of upstream splicing regulators.    Conclusions  This study led to uncover the molecular evolution of  Aedes aegypti dsx  splicing regulation with the respect of the more closely related Culicidae  Anopheles gambiae  orthologue. In  Aedes aegypti , the  dsx  gene is sex-specifically regulated and encodes two female-specific and one male-specific isoforms, all sharing a  doublesex / mab-3  (DM) domain-containing N-terminus and different C-termini. The sex-specific regulation is based on a combination of exon skipping, 5' alternative splice site choice and, most likely, alternative polyadenylation. Interestingly, when the  Aeadsx  gene is compared to the  Anopheles dsx  ortholog, there are differences in the  in silico  predicted default and regulated sex-specific splicing events, which suggests that the upstream regulators either are different or act in a slightly different manner. Furthermore, this study is a premise for the future development of transgenic sexing strains in mosquitoes useful for sterile insect technique (SIT) programs.

bmc evolutionary biology

Genomic organization and splicing evolution of the doublesex gene, a Drosophila regulator of sexual differentiation, in the dengue and yellow fever mosquito Aedes aegypti