Loss of YY1 Impacts the Heterochromatic State and Meiotic Double-Strand Breaks during Mouse Spermatogenesis

ABSTRACT The progression of spermatogenesis involves global changes in chromatin structure and conformation. However, our understanding of the regulation of chromatin changes in germ cells remains limited. Here we describe both in vivo RNA interference and genetic mouse knockout studies that identify a critical role for Yin Yang 1 (YY1) in mammalian spermatogenesis. In the YY1-deficient spermatocytes, we find a significant decrease in the global level of the heterochromatin markers (H3K9me3 and HP1-gamma) and a concomitant increase in the double-strand break (DSB) signals on chromosomes (gamma-H2AX, terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling, and Rad51) at the leptotene/zygotene stages of spermatocytes. These findings support a link between chromatin modifications and meiotic DSB formation, as has been seen in other model organisms. We propose that a depletion of YY1 may alter the structural integrity of heterochromatin, rendering it more accessible to the DSB machinery. In addition, YY1-deficient spermatocytes show univalent formation, increased aneuploidy, and pachytene cell death, which are likely due to defects in DNA repair. Taken together, this study identifies an important role for YY1 in mouse meiosis and provides new insight into mechanisms that regulate mammalian spermatogenesis.

[1]  G. Benvenuto,et al.  The INO80 protein controls homologous recombination in Arabidopsis thaliana. , 2004, Molecular cell.

[2]  V. Pirrotta,et al.  Drosophila Enhancer of Zeste/ESC Complexes Have a Histone H3 Methyltransferase Activity that Marks Chromosomal Polycomb Sites , 2002, Cell.

[3]  Z. Darżynkiewicz,et al.  Detection of DNA strand breaks in individual apoptotic cells by the in situ terminal deoxynucleotidyl transferase and nick translation assays. , 1993, Cancer research.

[4]  S. Ben‐Sasson,et al.  Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation , 1992, The Journal of cell biology.

[5]  N. Nakatsuji,et al.  RNA interference during spermatogenesis in mice. , 2005, Developmental biology.

[6]  L. Riou,et al.  Flow cytometric characterization of viable meiotic and postmeiotic cells by Hoechst 33342 in mouse spermatogenesis , 2005, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[7]  G. Schatten,et al.  Contractile apparatus of the normal and abortive cytokinetic cells during mouse male meiosis. , 2000, Journal of cell science.

[8]  P. Mieczkowski,et al.  The histone methylase Set2p and the histone deacetylase Rpd3p repress meiotic recombination at the HIS4 meiotic recombination hotspot in Saccharomyces cerevisiae. , 2008, DNA repair.

[9]  Malgorzata Schelder,et al.  A Polycomb group protein complex with sequence-specific DNA-binding and selective methyl-lysine-binding activities. , 2006, Genes & development.

[10]  R. Hawley,et al.  Chromosome Choreography: The Meiotic Ballet , 2003, Science.

[11]  N. Kleckner,et al.  The Single-End Invasion An Asymmetric Intermediate at the Double-Strand Break to Double-Holliday Junction Transition of Meiotic Recombination , 2001, Cell.

[12]  K. Rajewsky,et al.  Yin Yang 1 is a critical regulator of B-cell development. , 2007, Genes & development.

[13]  Y. Shi,et al.  Transcription factor Yin Yang 1 stimulates poly(ADP-ribosyl)ation and DNA repair. , 2001, Biochemical and biophysical research communications.

[14]  L. Anderson,et al.  Distribution of crossing over on mouse synaptonemal complexes using immunofluorescent localization of MLH1 protein. , 1999, Genetics.

[15]  J. Martens,et al.  Partitioning and plasticity of repressive histone methylation states in mammalian chromatin. , 2003, Molecular cell.

[16]  M. Washburn,et al.  YY1 functions with INO80 to activate transcription , 2007, Nature Structural &Molecular Biology.

[17]  J. Suja,et al.  Squash Procedure for Protein Immunolocalization in Meiotic Cells , 1998, Chromosome Research.

[18]  M. Hoekstra,et al.  Changes in protein composition of meiotic nodules during mammalian meiosis. , 1998, Journal of cell science.

[19]  James Allan,et al.  Formation of facultative heterochromatin in the absence of HP1 , 2003, The EMBO journal.

[20]  J. Ericsson,et al.  YY1 inhibits the activation of the p53 tumor suppressor in response to genotoxic stress. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[21]  Jeng-Shin Lee,et al.  Everything you have ever wanted to know about Yin Yang 1...... , 1997, Biochimica et biophysica acta.

[22]  E. Revenkova,et al.  Meiotic cohesin REC8 marks the axial elements of rat synaptonemal complexes before cohesins SMC1β and SMC3 , 2003, The Journal of cell biology.

[23]  U. Eichenlaub-Ritter.  Parental age‐related aneuploidy in human germ cells and offspring: A story of past and present , 1996, Environmental and molecular mutagenesis.

[24]  T. Ashley,et al.  Presynaptic association of Rad51 protein with selected sites in meiotic chromatin. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[25]  K. Nave,et al.  The Transcription Factor Yin Yang 1 Is Essential for Oligodendrocyte Progenitor Differentiation , 2007, Neuron.

[26]  N. Rajewsky,et al.  Survival of Resting Mature B Lymphocytes Depends on BCR Signaling via the Igα/β Heterodimer , 2004, Cell.

[27]  G. Almouzni,et al.  HP1 and the dynamics of heterochromatin maintenance , 2004, Nature Reviews Molecular Cell Biology.

[28]  R H Borts,et al.  Mlh1 is unique among mismatch repair proteins in its ability to promote crossing-over during meiosis. , 1997, Genes & development.

[29]  O. Ohara,et al.  Mammalian Polycomb Scmh1 mediates exclusion of Polycomb complexes from the XY body in the pachytene spermatocytes , 2006, Development.

[30]  J. Szostak,et al.  Double-strand breaks at an initiation site for meiotic gene conversion , 1989, Nature.

[31]  Yang Shi,et al.  Essential Dosage-Dependent Functions of the Transcription Factor Yin Yang 1 in Late Embryonic Development and Cell Cycle Progression , 2006, Molecular and Cellular Biology.

[32]  Alain Nicolas,et al.  Histone H3 lysine 4 trimethylation marks meiotic recombination initiation sites , 2009, The EMBO journal.

[33]  F. Collins,et al.  Meiotic arrest and aneuploidy in MLH3-deficient mice , 2002, Nature Genetics.

[34]  David J. Chen,et al.  Rad51 immunocytology in rat and mouse spermatocytes and oocytes , 1997, Chromosoma.

[35]  K. Rajewsky,et al.  Vagaries of conditional gene targeting , 2007, Nature Immunology.

[36]  Hengbin Wang,et al.  Role of Histone H3 Lysine 27 Methylation in Polycomb-Group Silencing , 2002, Science.

[37]  Yang Shi,et al.  A YY1–INO80 complex regulates genomic stability through homologous recombination–based repair , 2007, Nature Structural &Molecular Biology.

[38]  A. Shinohara,et al.  Rad51 protein involved in repair and recombination in S. cerevisiae is a RecA-like protein , 1992, Cell.

[39]  N. Kleckner,et al.  A pathway for generation and processing of double-strand breaks during meiotic recombination in S. cerevisiae , 1990, Cell.

[40]  P. Park,et al.  Postmeiotic Sex Chromatin in the Male Germline of Mice , 2006, Current Biology.

[41]  M. Whiteley,et al.  The Drosophila Polycomb group gene pleiohomeotic encodes a DNA binding protein with homology to the transcription factor YY1. , 1998, Molecular cell.

[42]  C. Heyting,et al.  A coiled‐coil related protein specific for synapsed regions of meiotic prophase chromosomes. , 1992, The EMBO journal.

[43]  G. Enders,et al.  Developmentally regulated expression of a mouse germ cell nuclear antigen examined from embryonic day 11 to adult in male and female mice. , 1994, Developmental biology.

[44]  C. Fritsch,et al.  The Drosophila pho-like gene encodes a YY1-related DNA binding protein that is redundant with pleiohomeotic in homeotic gene silencing , 2003, Development.

[45]  M. Deol.  Genetic Variants and Strains of the Laboratory Mouse , 1983 .

[46]  M. Lyon,et al.  Genetic variants and strains of the laboratory mouse , 1989 .

[47]  Karl Mechtler,et al.  Loss of the Suv39h Histone Methyltransferases Impairs Mammalian Heterochromatin and Genome Stability , 2001, Cell.

[48]  Yang Shi,et al.  Gene silencing by a DNA vector-based RNAi technology. , 2005, Methods in molecular biology.

[49]  V. Orlando.  Polycomb, Epigenomes, and Control of Cell Identity , 2003, Cell.

[50]  Y. Shinkai,et al.  Functional dynamics of H3K9 methylation during meiotic prophase progression , 2007, The EMBO journal.

[51]  R. B. Redmon,et al.  Identity , 2021, Notre Dame J. Formal Log..

[52]  G. Boissonneault,et al.  Transient DNA Strand Breaks During Mouse and Human Spermiogenesis: New Insights in Stage Specificity and Link to Chromatin Remodeling1 , 2004, Biology of reproduction.

[53]  P. de Boer,et al.  SHORT COMMUNICATIONS A drying-down technique for the spreading of mammalian meiocytes from the male and female germline , 1997, Chromosome Research.

[54]  Jung-Ae Kim,et al.  Heterochromatin is refractory to γ-H2AX modification in yeast and mammals , 2007, The Journal of cell biology.

[55]  S. Keeney,et al.  Recombinational DNA double-strand breaks in mice precede synapsis , 2001, Nature Genetics.

[56]  Yang Shi,et al.  Targeted Disruption of Mouse Yin Yang 1 Transcription Factor Results in Peri-Implantation Lethality , 1999, Molecular and Cellular Biology.

[57]  M. T. Parra,et al.  Involvement of the cohesin Rad21 and SCP3 in monopolar attachment of sister kinetochores during mouse meiosis I , 2004, Journal of Cell Science.

[58]  J. Lieb,et al.  Loss of a histone deacetylase dramatically alters the genomic distribution of Spo11p-catalyzed DNA breaks in Saccharomyces cerevisiae , 2007, Proceedings of the National Academy of Sciences.

[59]  Y. Shi,et al.  Poly(ADP-ribosyl)ation of transcription factor Yin Yang 1 under conditions of DNA damage. , 2001, Biochemical and biophysical research communications.

[60]  D. Wolgemuth,et al.  Primary Spermatocyte-Specific Cre Recombinase Activity in Transgenic Mice , 2004, Transgenic research.

[61]  N. Bonini,et al.  Transcription factor YY1 functions as a PcG protein in vivo , 2003, The EMBO journal.

[62]  Ali Hamiche,et al.  A chromatin remodelling complex involved in transcription and DNA processing , 2000, Nature.

[63]  A. Villeneuve,et al.  C. elegans HIM-17 Links Chromatin Modification and Competence for Initiation of Meiotic Recombination , 2004, Cell.

[64]  A. Schleiffer,et al.  A Caenorhabditis elegans cohesion protein with functions in meiotic chromosome pairing and disjunction. , 2001, Genes & development.

[65]  Tom Misteli,et al.  Maintenance of Stable Heterochromatin Domains by Dynamic HP1 Binding , 2003, Science.