title: The splicing landscape is globally reprogrammed during male meiosis.
abstract: Meiosis requires conserved transcriptional changes, but it is not known whether there is a corresponding set of RNA splicing switches. Here, we used RNAseq of mouse testis to identify changes associated with the progression from mitotic spermatogonia to meiotic spermatocytes. We identified ∼150 splicing switches, most of which affect conserved protein-coding exons. The expression of many key splicing regulators changed in the course of meiosis, including downregulation of polypyrimidine tract binding protein (PTBP1) and heterogeneous nuclear RNP A1, and upregulation of nPTB, Tra2β, muscleblind, CELF proteins, Sam68 and T-STAR. The sequences near the regulated exons were significantly enriched in target sites for PTB, Tra2β and STAR proteins. Reporter minigene experiments investigating representative exons in transfected cells showed that PTB binding sites were critical for splicing of a cassette exon in the Ralgps2 mRNA and a shift in alternative 5' splice site usage in the Bptf mRNA. We speculate that nPTB might functionally replace PTBP1 during meiosis for some target exons, with changes in the expression of other splicing factors helping to establish meiotic splicing patterns. Our data suggest that there are substantial changes in the determinants and patterns of alternative splicing in the mitotic-to-meiotic transition of the germ cell cycle.
authors.affiliation: Department of Biochemistry, University of Leicester, Leicester, LE1 9HN, UK, Institute of Genetic Medicine, Newcastle University, Newcastle upon Tyne, NE1 3BZ, UK, School of Biological and Biomedical Sciences, Durham University, Durham, DH1 3LE, UK, Department of Health Sciences, University of 00135 Rome 'Foro Italico', Rome, Italy, Laboratories of Neuroembryology and of Cellular and Molecular Neurobiology, Fondazione Santa Lucia IRCCS, 00143 Rome, Italy, Department of Public Health and Cell Biology, University of Rome Tor Vergata, 00133 Rome, Italy, Institute of Particle Physics Phenomenology, Durham University, Durham, DH1 3LE, UK and Life Technologies Ltd., Paisley PA4 9RF, UK.
authors.forename: Ralf
authors.initials: R
authors.lastname: Schmid
authors.forename: Sushma Nagaraja
authors.initials: SN
authors.lastname: Grellscheid
authors.forename: Ingrid
authors.initials: I
authors.lastname: Ehrmann
authors.forename: Caroline
authors.initials: C
authors.lastname: Dalgliesh
authors.forename: Marina
authors.initials: M
authors.lastname: Danilenko
authors.forename: Maria Paola
authors.initials: MP
authors.lastname: Paronetto
authors.forename: Simona
authors.initials: S
authors.lastname: Pedrotti
authors.forename: David
authors.initials: D
authors.lastname: Grellscheid
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Dixon
authors.forename: Claudio
authors.initials: C
authors.lastname: Sette
authors.forename: Ian C
authors.initials: IC
authors.lastname: Eperon
authors.forename: David J
authors.initials: DJ
authors.lastname: Elliott
mesh_terms: D017398:Alternative Splicing; D000818:Animals; D001483:Base Sequence; D005091:Exons; D008297:Male; D008540:Meiosis / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D059368:RNA Isoforms / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D017423:Sequence Analysis, RNA; D013090:Spermatocytes / Q000378:metabolism; D013093:Spermatogonia / Q000378:metabolism; D013737:Testis / Q000378:metabolism*; D059467:Transcriptome
chemical_list: D059368:RNA Isoforms; D016601:RNA-Binding Proteins
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