Suchergebnisse

© 2019 The Author(s). ; Mammalian gametogenesis involves dramatic and tightly regulated chromatin remodeling, whose regulatory pathways remain largely unexplored. Here, we generate a comprehensive high-resolution structural and functional atlas of mouse spermatogenesis by combining in situ chromosome conformation capture sequencing (Hi-C), RNA sequencing (RNA-seq), and chromatin immunoprecipitation sequencing (ChIP-seq) of CCCTC-binding factor (CTCF) and meiotic cohesins, coupled with confocal and super-resolution microscopy. Spermatogonia presents well-defined compartment patterns and topological domains. However, chromosome occupancy and compartmentalization are highly re-arranged during prophase I, with cohesins bound to active promoters in DNA loops out of the chromosomal axes. Compartment patterns re-emerge in round spermatids, where cohesin occupancy correlates with transcriptional activity of key developmental genes. The compact sperm genome contains compartments with actively transcribed genes but no fine-scale topological domains, concomitant with the presence of protamines. Overall, we demonstrate how genome-wide cohesin occupancy and transcriptional activity is associated with three-dimensional (3D) remodeling during spermatogenesis, ultimately reprogramming the genome for the next generation. ; This work was supported by the Ministry of Economy and Competitiveness ( BFU2017-89408-R to A.M.P.; BFU2013-47736-P and BFU2017-85926-P to M.A.M.-R.; and CGL2014-54317-P and CGL2017-83802-P to A.R.-H.) and the Agència de Gestió d'Ajuts Universitaris i de Recerca (AGAUR) ( DI2015 to A.R.-H. and R.A.C., as well as SGR468 to M.A.M-R.). Work at CRG, BIST, and UPF was in part funded by the Spanish Ministry of Economy and Competitiveness , 'Centro de Excelencia Severo Ochoa 2013-2017' (SEV-2012-0208), and 'Centro de Excelencia María de Maeztu 2016-2019.' CIC-IBMCC is supported by the Programa de Apoyo a Planes Estratégicos de Investigación de Estructuras de Investigación de Excelencia , co-funded by Junta de Castilla y León ( CSI239P18 ) and the European Regional Development Fund ( CLC–2017–01 ). C.V. is supported by a FPI predoctoral fellowship from the Ministry of Economy and Competitiveness ( BES-2015-072924 ). A.P.-G. is supported by a 'Doctorats Industrials' predoctoral fellowship ( AGAUR ). H.H. is a Miguel Servet ( CP14/00229 ) researcher funded by the Spanish Institute of Health Carlos III (ISCIII), the Agencia Estatal de Investigación (AEI), and FEDER ( SAF2017-89109-P ). C.M. is an Asociación Española Contra el Cáncer (AECC) postdoctoral fellow. M.A.M.-R. acknowledges support by the European Research Council under the 7th Framework Program FP7/2007-2013 (ERC grant agreement 609989 ) and the European Union's Horizon 2020 Research and Innovation Programme (grant agreement 676556 ). A.M.P. and A.R.-H. also acknowledge support from MeioNet ( BFU2015-71786-REDT ).

The spatial folding of chromosomes inside the nucleus has regulatory effects on gene expression, yet the impact of genome reshuffling on this organization remains unclear. Here, we take advantage of chromosome conformation capture in combination with single-nucleotide polymorphism (SNP) genotyping and analysis of crossover events to study how the higher-order chromatin organization and recombination landscapes are affected by chromosomal fusions in the mammalian germ line. We demonstrate that chromosomal fusions alter the nuclear architecture during meiosis, including an increased rate of heterologous interactions in primary spermatocytes, and alterations in both chromosome synapsis and axis length. These disturbances in topology were associated with changes in genomic landscapes of recombination, resulting in detectable genomic footprints. Overall, we show that chromosomal fusions impact the dynamic genome topology of germ cells in two ways: (i) altering chromosomal nuclear occupancy and synapsis, and (ii) reshaping landscapes of recombination. ; This work was supported by the Ministry of Economy and Competitiveness (CGL2014-54317-P and CGL2017-83802-P to A.R.-H., BFU2013-47736-P and BFU2017-85926-P to M.A.M.-R. and CGL2010-15243 to J.V.) and the Agència de Gestió d'Ajuts Universitaris i de Recerca, AGAUR (DI2015 and SGR1215 to A.R-H. as well as SGR468 to M.A.M.-R.). C.V. and L.A.-G. were supported by FPI predoctoral fellowships from the Ministry of Economy and Competitiveness (BES-2015-072924 and PRE-2018-083257). L.M.-G. was supported by an FPU predoctoral fellowships from the Ministry of Science, Innovation and University (FPU18/03867). M.A.M.-R. acknowledges support by the European Research Council under the 7th Framework Program FP7/2007-2013 (ERC grant agreement 609989) and the European Union's Horizon 2020 research and innovation program (grant agreement 676556). A.R.-H. also acknowledges support from MeioNet (BFU2015-71786-REDT).