Male-Specific Fruitless Isoforms Target Neurodevelopmental Genes to Specify a Sexually Dimorphic Nervous System

Summary Background In Drosophila, male courtship behavior is regulated in large part by the gene fruitless (fru). fru encodes a set of putative transcription factors that promote male sexual behavior by controlling the development of sexually dimorphic neuronal circuitry. Little is known about how Fru proteins function at the level of transcriptional regulation or the role that isoform diversity plays in the formation of a male-specific nervous system. Results To characterize the roles of sex-specific Fru isoforms in specifying male behavior, we generated novel isoform-specific mutants and used a genomic approach to identify direct Fru isoform targets during development. We demonstrate that all Fru isoforms directly target genes involved in the development of the nervous system, with individual isoforms exhibiting unique binding specificities. We observe that fru behavioral phenotypes are specified by either a single isoform or a combination of isoforms. Finally, we illustrate the utility of these data for the identification of novel sexually dimorphic genomic enhancers and novel downstream regulators of male sexual behavior. Conclusions These findings suggest that Fru isoform diversity facilitates both redundancy and specificity in gene expression, and that the regulation of neuronal developmental genes may be the most ancient and conserved role of fru in the specification of a male-specific nervous system.

[1]  J. C. Hall,et al.  Spatial, temporal, and sexually dimorphic expression patterns of the fruitless gene in the Drosophila central nervous system. , 2000, Journal of neurobiology.

[2]  M. Koganezawa,et al.  Fruitless Recruits Two Antagonistic Chromatin Factors to Establish Single-Neuron Sexual Dimorphism , 2012, Cell.

[3]  E. Tauber,et al.  Acoustic communication in Drosophila , 2003, Behavioural Processes.

[4]  M. Lehmann,et al.  Pipsqueak and GAGA factor act in concert as partners at homeotic and many other loci , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[5]  Tzumin Lee,et al.  Gradients of the Drosophila Chinmo BTB-Zinc Finger Protein Govern Neuronal Temporal Identity , 2006, Cell.

[6]  Jeffrey C. Hall,et al.  Neurogenetics of courtship and mating in Drosophila. , 2008, Advances in genetics.

[7]  Y. Choffat,et al.  Sexual behaviour in Drosophila is irreversibly programmed during a critical period , 1998, Current Biology.

[8]  Hinton,et al.  The courtship song of African Drosophila melanogaster , 1999 .

[9]  L. Beukeboom,et al.  The Fruitless gene in Nasonia displays complex sex-specific splicing and contains new zinc finger domains. , 2009, Molecular biology and evolution.

[10]  Wolfgang Huber,et al.  Ringo – an R/Bioconductor package for analyzing ChIP-chip readouts , 2007, BMC Bioinformatics.

[11]  Hania J. Pavlou,et al.  Courtship behavior in Drosophila melanogaster: towards a ‘courtship connectome’ , 2013, Current Opinion in Neurobiology.

[12]  P T Barnes,et al.  Extended reproductive roles of the fruitless gene in Drosophila melanogaster revealed by behavioral analysis of new fru mutants. , 1997, Genetics.

[13]  Caleb Webber,et al.  GAT: a simulation framework for testing the association of genomic intervals , 2013, Bioinform..

[14]  M. Ritchie,et al.  Drosophila song as a species-specific mating signal and the behavioural importance of Kyriacou & Hall cycles in D.melanogaster song , 1999, Animal Behaviour.

[15]  Toshiro Aigaki,et al.  Alternative splicing of lola generates 19 transcription factors controlling axon guidance in Drosophila , 2003, Nature Neuroscience.

[16]  B. Beutler,et al.  The BTB-ZF transcription factors , 2012, Cell cycle.

[17]  Elizabeth J. Rideout,et al.  Control of Sexual Differentiation and Behavior by the doublesex gene in Drosophila melanogaster , 2010, Nature Neuroscience.

[18]  S. Aerts,et al.  i-cisTarget: an integrative genomics method for the prediction of regulatory features and cis-regulatory modules , 2012, Nucleic acids research.

[19]  Tom H. Pringle,et al.  The human genome browser at UCSC. , 2002, Genome research.

[20]  L. Restifo,et al.  Anciently duplicated Broad Complex exons have distinct temporal functions during tissue morphogenesis , 2007, Development Genes and Evolution.

[21]  D. Haussler,et al.  Evolutionarily conserved elements in vertebrate, insect, worm, and yeast genomes. , 2005, Genome research.

[22]  B. S. Baker,et al.  Molecular genetic dissection of the sex-specific and vital functions of the Drosophila melanogaster sex determination gene fruitless. , 2001, Genetics.

[23]  K. Ui-Tei,et al.  Formation of the male-specific muscle in female Drosophila by ectopic fruitless expression , 2000, Nature Cell Biology.

[24]  B. S. Baker,et al.  The fruitless gene is required for the proper formation of axonal tracts in the embryonic central nervous system of Drosophila. , 2002, Genetics.

[25]  B. S. Baker,et al.  Direct targets of the D. melanogaster DSXF protein and the evolution of sexual development , 2011, Development.

[26]  K. Vasquez,et al.  Gene Targeting by Homologous Recombination , 2014 .

[27]  S. Kidd,et al.  DSL-Notch Signaling in the Drosophila Brain in Response to Olfactory Stimulation , 2011, Neuron.

[28]  S. Henikoff,et al.  Distinct HP1 and Su(var)3-9 complexes bind to sets of developmentally coexpressed genes depending on chromosomal location. , 2003, Genes & development.

[29]  A. Brand,et al.  Neural stem cell transcriptional networks highlight genes essential for nervous system development , 2009, The EMBO journal.

[30]  C. Kyriacou,et al.  Genetic variability of the interpulse interval of courtship song among some European populations of Drosophila melanogaster , 1994, Heredity.

[31]  Jai Y. Yu,et al.  Cellular Organization of the Neural Circuit that Drives Drosophila Courtship Behavior , 2010, Current Biology.

[32]  T. Duke,et al.  Active Process Mediates Species-Specific Tuning of Drosophila Ears , 2011, Current Biology.

[33]  M. Koganezawa,et al.  Neuronal Synaptic Outputs Determine the Sexual Fate of Postsynaptic Targets , 2010, Current Biology.

[34]  Jeffrey C. Hall,et al.  Control of Male Sexual Behavior and Sexual Orientation in Drosophila by the fruitless Gene , 1996, Cell.

[35]  Tony D. Southall,et al.  Prospero acts as a binary switch between self-renewal and differentiation in Drosophila neural stem cells. , 2006, Developmental cell.

[36]  Daniel H. Huson,et al.  Dendroscope: An interactive viewer for large phylogenetic trees , 2007, BMC Bioinformatics.

[37]  M. Arbeitman,et al.  Genomic and Functional Studies of Drosophila Sex Hierarchy Regulated Gene Expression in Adult Head and Nervous System Tissues , 2007, PLoS genetics.

[38]  Matthew S. Lebo,et al.  Ecdysone Receptor Acts in fruitless- Expressing Neurons to Mediate Drosophila Courtship Behaviors , 2009, Current Biology.

[39]  J. Dow,et al.  Using FlyAtlas to identify better Drosophila melanogaster models of human disease , 2007, Nature Genetics.

[40]  J. Allendorfer,et al.  Isoform-Specific Control of Male Neuronal Differentiation and Behavior in Drosophila by the fruitless Gene , 2006, Current Biology.

[41]  J. Billeter,et al.  The Sex-Determination Genes fruitless and doublesex Specify a Neural Substrate Required for Courtship Song , 2007, Current Biology.

[42]  C. Chothia,et al.  Evolution of the Protein Repertoire , 2003, Science.

[43]  Chris P. Ponting,et al.  Rapid Turnover of Long Noncoding RNAs and the Evolution of Gene Expression , 2012, PLoS genetics.

[44]  Daisuke Yamamoto,et al.  Fruitless specifies sexually dimorphic neural circuitry in the Drosophila brain , 2005, Nature.

[45]  Y. Rong,et al.  Gene targeting by homologous recombination in Drosophila. , 2000, Science.

[46]  S. Henikoff,et al.  Identification of in vivo DNA targets of chromatin proteins using tethered Dam methyltransferase , 2000, Nature Biotechnology.

[47]  A. Gardiner,et al.  The evolution of novelty in conserved genes; evidence of positive selection in the Drosophila fruitless gene is localised to alternatively spliced exons , 2013, Heredity.

[48]  R. Heinrich,et al.  Drosophila female precopulatory behavior is modulated by ecdysteroids. , 2012, Journal of insect physiology.

[49]  D. Yamamoto,et al.  Sexual orientation in Drosophila is altered by the satori mutation in the sex-determination gene fruitless that encodes a zinc finger protein with a BTB domain. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[50]  B. Graveley Alternative splicing: increasing diversity in the proteomic world. , 2001, Trends in genetics : TIG.

[51]  M. Bate,et al.  Innervation is essential for the development and differentiation of a sex-specific adult muscle in Drosophila melanogaster. , 1995, Development.

[52]  Trey Ideker,et al.  Cytoscape 2.8: new features for data integration and network visualization , 2010, Bioinform..

[53]  D. Kvitsiani,et al.  Neural Circuitry that Governs Drosophila Male Courtship Behavior , 2005, Cell.

[54]  Julie H. Simpson,et al.  A GAL4-driver line resource for Drosophila neurobiology. , 2012, Cell reports.

[55]  D. Yamamoto,et al.  Sex-switching of the Drosophila brain by two antagonistic chromatin factors , 2013, Fly.