Evolution of the trans-splicing Drosophila locus mod(mdg4) in several species of Diptera and Lepidoptera.

The modifier of mdg4 (mod[mdg4]) locus of Drosophila melanogaster (Dme) encodes chromatin proteins which are involved in position effect variegation, establishment of chromatin boundaries, nerve pathfinding, meiotic chromosome pairing and apoptosis. It was recently shown that mRNA trans-splicing is involved in the generation of at least 26 different mod(mdg4) transcripts. Here, we show that a similar complex mod(mdg4) locus exists in Drosophila pseudoobscura (Dps), Drosophila virilis (Dvi), Anopheles gambiae (Aga) and Bombyx mori (Bmo). As in D. melanogaster, most isoforms of these species contain a strongly conserved BTB/POZ domain (hereafter referred to as BTB domain) within the common N-terminal part and a Cys(2)His(2) motif containing FLYWCH domain within the isoform-specific C-terminal parts. By sequence comparison, we identified six novel isoforms in D. melanogaster and show that altogether 31 isoforms are perfectly conserved by sequence and position in the mod(mdg4) locus of the Drosophila species analyzed. We found significant differences in evolutionary speed of synonymous/nonsynonymous divergence between the various isoform specific exons. These results were extended by tree reconstruction analysis based on the evolved FLYWCH domains of predicted Mod(mdg4) proteins in Drosophila and Anopheles. Comparative analysis of mod(mdg4) gene structure in species of dipterans implicates that several internal inversions occurred making the mRNA trans-splicing mechanism indispensable for mod(mdg4) expression. Finally, we propose a model for the evolution of trans-splicing implementing effective regulation of many alternative gene products in a composite gene structure.