Abstract
takeout (to) is one of the male-specific genes expressed in the fat body that regulate male courtship behavior, and has been shown to act as a secreted protein in conjunction with courtship circuits. There are 23 takeout family members in Drosophila melanogaster, and homologues of this family are distributed across insect species. Sequence conservation among family members is low. Here we test the functional conservation of takeout family members by examining whether they can rescue the takeout courtship defect. We find that despite their sequence divergence takeout members from Aedes aegypti and Epiphas postvittana, as well as family members from D. melanogaster can substitute for takeout in courtship, demonstrating their functional conservation. Making use of the known E. postvittana Takeout structure, we used homology modeling and amphipathic helix analysis and found high overall structural conservation, including high conservation of the structure and amphipathic lining of an internal cavity that has been shown to accommodate hydrophobic ligands. Together these data suggest a high degree of structural conservation that likely underlies functional conservation in courtship. In addition, we have identified a role for a conserved exposed protein motif important for the protein’s role in courtship.
Highlights
Courtship rituals in Drosophila melanogaster consist of a series of stereotyped behaviors displayed by the male in order to gain access to and mate with females [1, 2]
Mutant males are capable of all steps of courtship, but display them with reduced frequency [7]. takeout is male- expressed in the head fat body, from where it is secreted into the hemolymph and acts as a secreted protein [8]. takeout has the characteristics of small soluble proteins and is most similar to Juvenile Hormone Binding Proteins (JHBPs) from other insects
UAS-A. aegypti To, UAS-D.mel To, UAS-Ep.To, UAS-B. mori JHBP and UAS-D.mel To-mut were established during this study by PCR using the primers indicated
Summary
Courtship rituals in Drosophila melanogaster consist of a series of stereotyped behaviors displayed by the male in order to gain access to and mate with females [1, 2]. With the exception of two conserved motifs the sequence conservation between family members is fairly low, the most distant paralog being only 18% identical (CG16820) Except for takeout, their specific functions are unknown, several exhibit circadian regulated expression, and all contain signal sequences indicative of secreted proteins. Current data suggest that takeout is part of a large gene family found throughout insects with roles in metabolism, circadian behavior, aging, and male courtship behavior. We observed many instances of gene duplication and loss and evidence of positive selection in several lineages [13], consistent with the action of sexual selection on male- expressed genes These findings raise the possibility that the takeout gene family is a group of conserved proteins that may have maintained similar functional roles across species among at least some of its members. Takeout protein structure with the structure of a previously crystallized Takeout protein from Epiphyas postvittana [19, 21] and find high structural conservation as a possible unifying functional feature among family members
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