title: Relocation facilitates the acquisition of short cis-regulatory regions that drive the expression of retrogenes during spermatogenesis in Drosophila.
abstract: Retrogenes are functional processed copies of genes that originate via the retrotranscription of an mRNA intermediate and often exhibit testis-specific expression. Although this expression pattern appears to be favored by selection, the origin of such expression bias remains unexplained. Here, we study the regulation of two young testis-specific Drosophila retrogenes, Dntf-2r and Pros28.1A, using genetic transformation and the enhanced green fluorescent protein reporter gene in Drosophila melanogaster. We show that two different short (<24 bp) regions upstream of the transcription start sites (TSSs) act as testis-specific regulatory motifs in these genes. The Dntf-2r regulatory region is similar to the known β2 tubulin 14-bp testis motif (β2-tubulin gene upstream element 1 [β2-UE1]). Comparative sequence analyses reveal that this motif was already present before the Dntf-2r insertion and was likely driving the transcription of a noncoding RNA. We also show that the β2-UE1 occurs in the regulatory regions of other testis-specific retrogenes, and is functional in either orientation. In contrast, the Pros28.1A testes regulatory region in D. melanogaster appears to be novel. Only Pros28.1B, an older paralog of the Pros28.1 gene family, seems to carry a similar regulatory sequence. It is unclear how the Pros28.1A regulatory region was acquired in D. melanogaster, but it might have evolved de novo from within a region that may have been preprimed for testes expression. We conclude that relocation is critical for the evolutionary origin of male germline-specific cis-regulatory regions of retrogenes because expression depends on either the site of the retrogene insertion or the sequence changes close to the TSS thereafter. As a consequence we infer that positive selection will play a role in the evolution of these regulatory regions and can often act from the moment of the retrocopy insertion.
authors.affiliation: Department of Biology, University of Texas at Arlington.
authors.forename: Mehran
authors.initials: M
authors.lastname: Sorourian
authors.affiliation: Department of Biology, University of Texas at Arlington.
authors.forename: Mansi M
authors.initials: MM
authors.lastname: Kunte
authors.affiliation: Department of Biology, University of Texas at Arlington.
authors.forename: Susana
authors.initials: S
authors.lastname: Domingues
authors.affiliation: Center for Integrative Bioinformatics Vienna (CIBIV), Max F Perutz Laboratories, University of Vienna and Medical University of Vienna, Vienna, Austria.
authors.forename: Miguel
authors.initials: M
authors.lastname: Gallach
authors.affiliation: Department of Biology, University of Texas at Arlington.
authors.forename: Fulya
authors.initials: F
authors.lastname: Özdil
authors.affiliation: Department of Biology, University of Texas at Arlington.
authors.forename: Javier
authors.initials: J
authors.lastname: Río
authors.affiliation: Department of Biology, University of Texas at Arlington betran@uta.edu.
authors.forename: Esther
authors.initials: E
authors.lastname: Betrán
mesh_terms: D000818:Animals; D029721:Drosophila Proteins / Q000235:genetics*; D004331:Drosophila melanogaster / Q000235:genetics / Q000502:physiology*; D018507:Gene Expression Regulation, Developmental; D008297:Male; D029543:Nucleocytoplasmic Transport Proteins / Q000235:genetics*; D009928:Organ Specificity; D046988:Proteasome Endopeptidase Complex / Q000235:genetics*; D050436:Regulatory Elements, Transcriptional; D018626:Retroelements; D012641:Selection, Genetic; D013737:Testis / Q000254:growth & development*
chemical_list: D029721:Drosophila Proteins; C503609:Ntf-2 protein, Drosophila; D029543:Nucleocytoplasmic Transport Proteins; D018626:Retroelements; C000595809:Prosalpha4T1 protein, Drosophila; D046988:Proteasome Endopeptidase Complex
keywords: Drosophila; gene duplication; gene relocation; regulatory element; retrogene; testis expression