DOCSLIB.ORG

Studying Meiotic Cohesin in Somatic Cells Reveals That Rec8-Containing

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Studying Meiotic Cohesin in Somatic Cells Reveals That Rec8-Containing © 2018. Published by The Company of Biologists Ltd | Journal of Cell Science (2018) 131, jcs212100. doi:10.1242/jcs.212100 SHORT REPORT Studying meiotic cohesin in somatic cells reveals that Rec8- containing cohesin requires Stag3 to function and is regulated by Wapl and sororin Peter G. Wolf*, Alexander Cuba Ramos, Julia Kenzel, Brigitte Neumann and Olaf Stemmann ABSTRACT when phosphorylation inactivates sororin (Dreier et al., 2011; Hauf The DNA-embracing, ring-shaped multiprotein complex cohesin et al., 2005; Liu et al., 2013). A small pool of cohesin, however, is mediates sister chromatid cohesion and is stepwise displaced in resistant to this so-called prophase pathway signalling. At mitosis by Wapl and separase (also known as ESPL1) to facilitate centromeres, shugoshin 1 (Sgo1) recruits the protein phosphatase anaphase. Proper regulation of chromosome cohesion throughout 2A (PP2A) to keep sororin active by means of dephosphorylation meiosis is critical for preventing formation of aneuploid gametes, (Liu et al., 2013; McGuinness et al., 2005; Tang et al., 2006). which are associated with trisomies and infertility in humans. Centromeric cohesion is resolved in anaphase when separase (also Studying cohesion in meiocytes is complicated by their difficult known as ESPL1) cleaves Scc1 from the remaining chromosomal experimental amenability and the absence of cohesin turnover. Here, cohesin (Hauf et al., 2001). we use cultured somatic cells to unravel fundamental aspects of In a prototypic meiosis, recombination events between non-sister meiotic cohesin. When expressed in Hek293 cells, the kleisin Rec8 chromatids create bivalents, in which the homologous displays no affinity for the peripheral cohesin subunits Stag1 or Stag2 chromosomes are held together by cohesin on chromosome arms and remains cytoplasmic. However, co-expression of Stag3 is located distally to crossovers. Segregation of homologous sufficient for Rec8 to enter the nucleus, load onto chromatin, and chromosomes in meiosis I is achieved by selective loss of cohesin functionally replace its mitotic counterpart Scc1 (also known as at chromosome arms. In the second meiotic division, cohesion is RAD21) during sister chromatid cohesion and dissolution. Rec8– resolved at the centromeres allowing the disjunction of sister Stag3 cohesin physically interacts with Pds5, Wapl and sororin (also chromatids. Interestingly, both waves of cohesin removal depend on known as CDCA5). Importantly, Rec8–Stag3 cohesin is shown to be separase activity. However, centromeric cohesion is maintained susceptible to Wapl-dependent ring opening and sororin-mediated during meiosis I because (1) Scc1 is functionally replaced by Rec8, protection. These findings exemplify that our model system is suitable which is different from its mitotic counterpart in that it must be to rapidly generate testable predictions for important unresolved phosphorylated in order to be cleaved by separase (Kudo et al., issues of meiotic cohesion regulation. 2009; Riedel et al., 2006) and (2) Sgo2, which replaces Sgo1 in mammalian meiosis, recruits PP2A to keep centromeric Rec8 in a KEY WORDS: Meiosis, Cohesin, Rec8, Stag3, Wapl, Sororin dephosphorylated state (Lee et al., 2008; Llano et al., 2008). An unresolved question is whether Sgo1 is only competent to INTRODUCTION protect Scc1 cohesin from the prophase pathway and whether, Cohesin is a multiprotein complex that provides sister chromatid conversely, Sgo2 might only shield Rec8 cohesin from separase. cohesion by embracing both DNA copies in its middle. In In addition to Stag1 and Stag2, germ cells contain a third variant mitotically dividing cells, an Smc1α–Smc3 heterodimer named Stag3 (Prieto et al., 2001). Furthermore, vertebrate meiocytes associates with the kleisin subunit Scc1 (also known as RAD21) express two variants of Smc1, Smc1α and the meiosis-specific forming a tripartite ring. Human somatic cells express two Smc1β (Revenkova et al., 2001; Revenkova et al., 2004). Which of variants of a fourth subunit, Stag1 and Stag2, which bind Scc1 in the possible combinations of cohesin subunits actually exist in germ a mutually exclusive manner (Nasmyth, 2011; Nasmyth and cells is controversial (Ishiguro et al., 2011; Lee and Hirano, 2011; Haering, 2009). Revenkova et al., 2004). Moreover, while it is clear that Rec8 is Vertebrate cohesin is loaded in telophase but is constantly crucial for cohesion in meiocytes (Tachibana-Konwalski et al., 2010), removed from chromatin owing to the action of Wapl (Gandhi et al., whether and which other meiosis-specific cohesin subunits are 2006; Kueng et al., 2006), until S phase when a pool of cohesin is essential for cohesion still needs clarification. Wapl is expressed stabilised by the Wapl antagonist sororin (also known as CDCA5) during meiosis, and mutant spermatocytes with dysregulated Wapl (Nishiyama et al., 2010; Rankin et al., 2005). In early mitosis, Wapl- exhibit increased chromosomal Smc3 levels (Brieño-Enríquez et al., dependent removal of cohesin from chromosome arms is boosted 2016; Kuroda et al., 2005). However, whether the Rec8 cohesin is responsive to Wapl-dependent ring opening remains enigmatic. Here, we exploit somatic cells to study human meiotic cohesin Chair of Genetics, University of Bayreuth, Universitätsstraße 30, 95440 Bayreuth, subunits and reveal the following: (1) Rec8 does not interact with Germany. Stag1 or Stag2 but robustly associates with Stag3 and becomes *Author for correspondence ([email protected]) functional (i.e. loaded onto chromatin and cohesive) only in presence of this meiosis-specific cohesin subunit; (2) Rec8 cohesin P.G.W., 0000-0002-2273-676X; A.C., 0000-0002-3630-1011; J.K., 0000-0002- 1094-3876; B.N., 0000-0002-2621-3851 physically interacts with and is regulated by sororin and its antagonist Wapl; (3) Sgo1 and Sgo2 are indeed specific for Received 17 October 2017; Accepted 26 April 2018 distinct cohesin complexes as specified by the kleisin subunit. Journal of Cell Science 1 SHORT REPORT Journal of Cell Science (2018) 131, jcs212100. doi:10.1242/jcs.212100 RESULTS AND DISCUSSION We generated transgenic Hek293 cell lines inducibly expressing C-terminally GFP-tagged Rec8 or, as a control, Scc1 and compared the cellular localisation of both fusion proteins. Notably, in all clones analysed the levels of Scc1–GFP were lower than Rec8–GFP (data not shown) consistent with previous experiments indicating that cells try to keep a constant level of Scc1 (Schöckel et al., 2011). Consistent with a recent report (Rong et al., 2017), we found that Rec8, in contrast to Scc1, was excluded from the nucleus (Fig. 1A). We reasoned that another meiosis-specific cohesin subunit might be necessary for the nuclear targeting of Rec8. First, we asked whether Rec8 is competent to associate with all non-kleisin subunits of the somatic cohesin complex. Immunoprecipitation (IP) experiments revealed a robust interaction of Rec8 with Smc3 and Smc1α (Fig. 1B). Interestingly, studies conducted in spermatocytes revealed an association of Rec8 only with Smc1β and not with Smc1α (Ishiguro et al., 2011; Lee and Hirano, 2011). Importantly, endogenous Stag1 and Stag2, while readily co-purifying with Scc1, failed to co-IP with Rec8 (Fig. 1C; Fig. S1A). We directly compared transiently expressed Flag-tagged Stag2 and Stag3 in their ability to associate with Rec8 and confirmed that only Stag3, and not Stag2, exhibited a strong interaction with the kleisin (Fig. 2A). Moreover, Stag3 changed the localisation of Rec8 from the cytoplasm to the nucleus, while ectopic expression of Stag2 had no such effect (Fig. 2B–D; Fig. S2A,B). In addition, the nuclear Rec8–GFP signal became resistant to preextraction in presence of Stag3, indicating that Rec8 is also loaded onto chromatin in a Stag3-dependent manner. Stag3 triggered this effect irrespectively of its expression from a transiently transfected plasmid (Fig. S2A,B) or a genomically integrated transgene (Fig. 2B–D). Rec8 behaved the same in these kinds of experiments with both a C-terminal GFP tag and an N-terminal Myc5 tag indicating that the tag did not affect kleisin localisation (Fig. S2C,D). Taken together, our data suggest that Rec8 might only be functional when assembled in a cohesin complex that also contains Stag3. Although some somatic cell types express Smc1β (Mannini et al., 2015), we could not detect any Smc1β transcript as judged by means of a PCR on isolated cDNA (Fig. S1B). Rigorous testing of whether Rec8-containing cohesin can mediate cohesion in somatic cells requires abrogation of Scc1 cohesin function. In our hands, the knockdown of SCC1 was incomplete and did not result in a penetrant phenotype. Therefore, we used overexpression of hyperactive separase (Boos et al., 2008; Holland and Taylor, 2006) or depletion of Sgo1 (McGuinness et al., 2005; see further details below) as two independent and established methods to induce premature sister chromatid separation (SCS) in prometaphase cells. We then asked whether the cohesion defect could be rescued by presence of Stag3 and Rec8. First, parental Hek293 cells, cells expressing Rec8 with Stag2, and Rec8 with Stag3 were transfected to express hyperactive separase and arrested with nocodazole treatment. Subsequent chromosome spreading revealed untimely SCS in an average of 31% of the cells without the transgene or with REC8 and STAG2 (Fig. 3A; lanes 2 and 3; Fig. S3A), which represents a fivefold increase relative to control cells (lane 1). Importantly, precocious Fig. 1. Rec8 interacts with
Load more

Recommended publications
  • Meiotic Cohesin and Variants Associated with Human Reproductive Aging and Disease
    fcell-09-710033 July 27, 2021 Time: 16:27 # 1 REVIEW published: 02 August 2021 doi: 10.3389/fcell.2021.710033 Meiotic Cohesin and Variants Associated With Human Reproductive Aging and Disease Rachel Beverley1, Meredith L. Snook1 and Miguel Angel Brieño-Enríquez2* 1 Division of Reproductive Endocrinology and Infertility, Department of Obstetrics, Gynecology, and Reproductive Sciences, University of Pittsburgh, Pittsburgh, PA, United States, 2 Magee-Womens Research Institute, Department of Obstetrics, Gynecology, and Reproductive Sciences, University of Pittsburgh, Pittsburgh, PA, United States Successful human reproduction relies on the well-orchestrated development of competent gametes through the process of meiosis. The loading of cohesin, a multi- protein complex, is a key event in the initiation of mammalian meiosis. Establishment of sister chromatid cohesion via cohesin rings is essential for ensuring homologous recombination-mediated DNA repair and future proper chromosome segregation. Cohesin proteins loaded during female fetal life are not replenished over time, and therefore are a potential etiology of age-related aneuploidy in oocytes resulting in Edited by: decreased fecundity and increased infertility and miscarriage rates with advancing Karen Schindler, Rutgers, The State University maternal age. Herein, we provide a brief overview of meiotic cohesin and summarize of New Jersey, United States the human genetic studies which have identified genetic variants of cohesin proteins and Reviewed by: the associated reproductive phenotypes
    [Show full text]
  • Atrec8 and Atscc3 Are Essential to the Monopolar Orientation of the Kinetochores During Meiosis
    Research Article 4621 AtREC8 and AtSCC3 are essential to the monopolar orientation of the kinetochores during meiosis Liudmila Chelysheva, Stéphanie Diallo*, Daniel Vezon, Ghislaine Gendrot, Nathalie Vrielynck, Katia Belcram, Nathalie Rocques, Angustias Márquez-Lema‡, Anuj M. Bhatt§, Christine Horlow, Raphaël Mercier, Christine Mézard and Mathilde Grelon¶ Institut Jean-Pierre Bourgin, Station de Génétique et d’Amélioration des Plantes, INRA de Versailles, Route de Saint-Cyr, 78026 Versailles CEDEX, France *Present address: Laboratoire de Microbiologie du Froid UPRES 2123, 55 Rue Saint-Germain, 27000 Evreux, France ‡Present address: Instituto de Agricultura Sostenible (CSIC), Apartado 4084, E-14080, Córdoba, Spain §Present address: Department of Plant Sciences, University of Oxford, South Parks Road, Oxford, OX1 3RB, UK ¶Author for correspondence (e-mail: [email protected]) Accepted 13 July 2005 Journal of Cell Science 118, 4621-4632 Published by The Company of Biologists 2005 doi:10.1242/jcs.02583 Summary The success of the first meiotic division relies (among other in meiotic nuclei as early as interphase, and bound to the factors) on the formation of bivalents between homologous chromosome axis from early leptotene through to anaphase chromosomes, the monopolar orientation of the sister I. We show here that both AtREC8 and AtSCC3 are kinetochores at metaphase I and the maintenance of necessary not only to maintain centromere cohesion at centromeric cohesion until the onset of anaphase II. The anaphase I, but also for the monopolar orientation of the meiotic cohesin subunit, Rec8 has been reported to be one kinetochores during the first meiotic division. We also of the key players in these processes, but its precise role in found that AtREC8 is involved in chromosome axis kinetochore orientation is still under debate.
    [Show full text]
  • Evolution, Expression and Meiotic Behavior of Genes Involved in Chromosome Segregation of Monotremes
    G C A T T A C G G C A T genes Article Evolution, Expression and Meiotic Behavior of Genes Involved in Chromosome Segregation of Monotremes Filip Pajpach , Linda Shearwin-Whyatt and Frank Grützner * School of Biological Sciences, The University of Adelaide, Adelaide, SA 5005, Australia; fi[email protected] (F.P.); [email protected] (L.S.-W.) * Correspondence: [email protected] Abstract: Chromosome segregation at mitosis and meiosis is a highly dynamic and tightly regulated process that involves a large number of components. Due to the fundamental nature of chromosome segregation, many genes involved in this process are evolutionarily highly conserved, but duplica- tions and functional diversification has occurred in various lineages. In order to better understand the evolution of genes involved in chromosome segregation in mammals, we analyzed some of the key components in the basal mammalian lineage of egg-laying mammals. The chromosome passenger complex is a multiprotein complex central to chromosome segregation during both mitosis and meio- sis. It consists of survivin, borealin, inner centromere protein, and Aurora kinase B or C. We confirm the absence of Aurora kinase C in marsupials and show its absence in both platypus and echidna, which supports the current model of the evolution of Aurora kinases. High expression of AURKBC, an ancestor of AURKB and AURKC present in monotremes, suggests that this gene is performing all necessary meiotic functions in monotremes. Other genes of the chromosome passenger complex complex are present and conserved in monotremes, suggesting that their function has been preserved Citation: Pajpach, F.; in mammals.
    [Show full text]
  • Redundant and Specific Roles of Cohesin STAG Subunits in Chromatin Looping and Transcriptional Control
    Downloaded from genome.cshlp.org on October 10, 2021 - Published by Cold Spring Harbor Laboratory Press Research Redundant and specific roles of cohesin STAG subunits in chromatin looping and transcriptional control Valentina Casa,1,6 Macarena Moronta Gines,1,6 Eduardo Gade Gusmao,2,3,6 Johan A. Slotman,4 Anne Zirkel,2 Natasa Josipovic,2,3 Edwin Oole,5 Wilfred F.J. van IJcken,1,5 Adriaan B. Houtsmuller,4 Argyris Papantonis,2,3 and Kerstin S. Wendt1 1Department of Cell Biology, Erasmus MC, 3015 GD Rotterdam, The Netherlands; 2Center for Molecular Medicine Cologne, University of Cologne, 50931 Cologne, Germany; 3Institute of Pathology, University Medical Center, Georg-August University of Göttingen, 37075 Göttingen, Germany; 4Optical Imaging Centre, Erasmus MC, 3015 GD Rotterdam, The Netherlands; 5Center for Biomics, Erasmus MC, 3015 GD Rotterdam, The Netherlands Cohesin is a ring-shaped multiprotein complex that is crucial for 3D genome organization and transcriptional regulation during differentiation and development. It also confers sister chromatid cohesion and facilitates DNA damage repair. Besides its core subunits SMC3, SMC1A, and RAD21, cohesin in somatic cells contains one of two orthologous STAG sub- units, STAG1 or STAG2. How these variable subunits affect the function of the cohesin complex is still unclear. STAG1- and STAG2-cohesin were initially proposed to organize cohesion at telomeres and centromeres, respectively. Here, we uncover redundant and specific roles of STAG1 and STAG2 in gene regulation and chromatin looping using HCT116 cells with an auxin-inducible degron (AID) tag fused to either STAG1 or STAG2. Following rapid depletion of either subunit, we perform high-resolution Hi-C, gene expression, and sequential ChIP studies to show that STAG1 and STAG2 do not co-occupy in- dividual binding sites and have distinct ways by which they affect looping and gene expression.
    [Show full text]
  • STAG3 Is a Strong Candidate Gene for Male Infertility
    Human Molecular Genetics, 2014, Vol. 23, No. 13 3421–3431 doi:10.1093/hmg/ddu051 Advance Access published on March 7, 2014 STAG3 is a strong candidate gene for male infertility Elena Llano1,∗,{, Laura Gomez-H3,{, Ignacio Garcı´a-Tun˜ o´ n3, Manuel Sa´nchez-Martı´n2, Sandrine Caburet4,5, Jose Luis Barbero6, John C. Schimenti7, Reiner A. Veitia4,5 and Alberto M. Pendas3,∗ Downloaded from https://academic.oup.com/hmg/article/23/13/3421/659338 by Universidad de Salamanca user on 01 September 2020 1Departamento de Fisiologı´a y Farmacologı´a and 2Departamento de Medicina, Universidad de Salamanca, 37007 Salamanca, Spain, 3Instituto de Biologı´a Molecular y Celular del Ca´ncer (CSIC-USAL), 37007 Salamanca, Spain, 4Institut Jacques Monod, Universite´ Paris Diderot, CNRS UMR7592, Paris 75013, France, 5Universite´ Paris Diderot-Paris 7, 75205 Paris Cedex 13, France, 6Centro de Investigaciones Biolo´gicas (CSIC), Madrid 28040, Spain and 7Center for Vertebrate Genomics, Cornell University, Ithaca, NY 14850, USA Received December 20, 2013; Revised and Accepted January 31, 2014 Oligo- and azoospermia are severe forms of male infertility. However, known genetic factors account only for a small fraction of the cases. Recently, whole-exome sequencing in a large consanguineous family with inherited premature ovarian failure (POF) identified a homozygous frameshift mutation in the STAG3 gene leading to a pre- mature stopcodon. STAG3 encodesa meiosis-specific subunit of the cohesincomplex, a large proteinaceous ring withDNA-entrappingabilitythatensures sister chromatid cohesionandenablescorrect synapsisandsegregation of homologous chromosomes during meiosis. The pathogenicity of the STAG3 mutations was functionally vali- datedwithaloss-of-functionmousemodelfor STAG3inoogenesis. However,andsince noneof the malemembers of this family was homozygous for the mutant allele, we only could hypothesized its putative involvement in male infertility.
    [Show full text]
  • Loss of Cohesin Complex Components STAG2 Or STAG3 Confers Resistance to BRAF Inhibition in Melanoma
    Loss of cohesin complex components STAG2 or STAG3 confers resistance to BRAF inhibition in melanoma The Harvard community has made this article openly available. Please share how this access benefits you. Your story matters Citation Shen, C., S. H. Kim, S. Trousil, D. T. Frederick, A. Piris, P. Yuan, L. Cai, et al. 2016. “Loss of cohesin complex components STAG2 or STAG3 confers resistance to BRAF inhibition in melanoma.” Nature medicine 22 (9): 1056-1061. doi:10.1038/nm.4155. http:// dx.doi.org/10.1038/nm.4155. Published Version doi:10.1038/nm.4155 Citable link http://nrs.harvard.edu/urn-3:HUL.InstRepos:31731818 Terms of Use This article was downloaded from Harvard University’s DASH repository, and is made available under the terms and conditions applicable to Other Posted Material, as set forth at http:// nrs.harvard.edu/urn-3:HUL.InstRepos:dash.current.terms-of- use#LAA HHS Public Access Author manuscript Author ManuscriptAuthor Manuscript Author Nat Med Manuscript Author . Author manuscript; Manuscript Author available in PMC 2017 March 01. Published in final edited form as: Nat Med. 2016 September ; 22(9): 1056–1061. doi:10.1038/nm.4155. Loss of cohesin complex components STAG2 or STAG3 confers resistance to BRAF inhibition in melanoma Che-Hung Shen1, Sun Hye Kim1, Sebastian Trousil1, Dennie T. Frederick2, Adriano Piris3, Ping Yuan1, Li Cai1, Lei Gu4, Man Li1, Jung Hyun Lee1, Devarati Mitra1, David E. Fisher1,2, Ryan J. Sullivan2, Keith T. Flaherty2, and Bin Zheng1,* 1Cutaneous Biology Research Center, Massachusetts General Hospital and Harvard Medical School, Charlestown, MA 2Department of Medical Oncology, Massachusetts General Hospital Cancer Center, Boston, MA 3Department of Dermatology, Brigham & Women's Hospital and Harvard Medical School, Boston, MA 4Division of Newborn Medicine, Boston Children's Hospital, Harvard Medical School, Boston, MA.
    [Show full text]
  • Low Tolerance for Transcriptional Variation at Cohesin Genes Is Accompanied by Functional Links to Disease-Relevant Pathways
    bioRxiv preprint doi: https://doi.org/10.1101/2020.04.11.037358; this version posted April 13, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. Title Low tolerance for transcriptional variation at cohesin genes is accompanied by functional links to disease-relevant pathways Authors William Schierdingǂ1, Julia Horsfieldǂ2,3, Justin O’Sullivan1,3,4 ǂTo whom correspondence should be addressed. 1 Liggins Institute, The University of Auckland, Auckland, New Zealand 2 Department of Pathology, Dunedin School of Medicine, University of Otago, Dunedin, New Zealand 3 The Maurice Wilkins Centre for Biodiscovery, The University of Auckland, Auckland, New Zealand 4 MRC Lifecourse Epidemiology Unit, University of Southampton Acknowledgements This work was supported by a Royal Society of New Zealand Marsden Grant to JH and JOS (16-UOO- 072), and WS was supported by the same grant. Contributions WS planned the study, performed analyses, and drafted the manuscript. JH and JOS revised the manuscript. Competing interests None declared. bioRxiv preprint doi: https://doi.org/10.1101/2020.04.11.037358; this version posted April 13, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. Abstract Variants in DNA regulatory elements can alter the regulation of distant genes through spatial- regulatory connections.
    [Show full text]
  • Characterization of the Major Human STAG3 Variants Using Some Proteomics and Bioinformatics Assays Inam J
    Lafta et al. Egyptian Journal of Medical Human Genetics (2020) 21:9 Egyptian Journal of Medical https://doi.org/10.1186/s43042-020-0051-0 Human Genetics RESEARCH Open Access Characterization of the major human STAG3 variants using some proteomics and bioinformatics assays Inam J. Lafta1*, Bassam K. Kudhair2 and Noralhuda N. Alabid3 Abstract Background: STAG3 is the meiotic component of cohesin and a member of the Cancer Testis Antigen (CTA) family. This gene has been found to be overexpressed in many types of cancer, and recently, its variants have been implicated in other disorders and many human diseases. Therefore, this study aimed to analyze the major variants of STAG3. Western blot (WB) and immunoprecipitation (IP) assays were performed using two different anti-STAG3 antibodies that targeted the relevant protein in MCF-7, T-47D, MDA-MB-468, and MDA-MB-231 breast cancer cells with Jurkat and MCF-10A cells as positive and negative controls, respectively. In silico analyses were searched to study the major isoforms. Results: WB and IP assays revealed two abundant polypeptides < 191 kDa and ~ 75 kDa in size. Specific bioinformatics tools successfully determined the three-dimensional (3-D) structure, the subcellular localization, and the secondary structures of the isoforms. Furthermore, some of the physicochemical properties of the STAG3 proteins were also determined. Conclusions: The results of this study revealed the power of applying the biological techniques (WB and IP) with the bioinformatics assays and the possibility of their exploitation in understanding diseased genes. Exploring the major variants of STAG3 at the protein level could help decipher some disorders associated with their occurrence, along with designing drugs effective at least for some relevant diseases.
    [Show full text]
  • STAG3 Homozygous Missense Variant Causes Primary Ovarian Insufficiency and Male Non- Obstructive Azoospermia
    STAG3 homozygous missense variant causes primary ovarian insufficiency and male non- obstructive azoospermia Running title: STAG3 variants in female and male infertility Downloaded from https://academic.oup.com/molehr/article-abstract/doi/10.1093/molehr/gaaa050/5868415 by Central Michigan University user on 09 July 2020 Sylvie Jaillard1,2,3, Kenneth McElreavy4, Gorjana Robevska1, Linda Akloul5, Farah Ghieh6, Rajini Sreenivasan1, Marion Beaumont3, Anu Bashamboo4, Joelle Bignon-Topalovic4, Anne-Sophie Neyroud8, Katrina Bell1, Elisabeth Veron-Gastard8, Erika Launay3, Jocelyn van den Bergen1, Bénédicte Nouyou3, François Vialard6,7, Marc-Antoine Belaud-Rotureau2,3,8, Katie L Ayers1,9, Sylvie Odent5, Célia Ravel2,8, Elena J Tucker1,9*, Andrew H Sinclair1,9* 1- Murdoch Children’s Research Institute, Royal Children’s Hospital, Melbourne, VIC Australia 2- Univ Rennes, CHU Rennes, INSERM, EHESP, IRSET (Institut de recherche en santé, environnement et travail) – UMR_S 1085, F-35000 Rennes, France 3- CHU Rennes, Service de Cytogénétique et Biologie Cellulaire, F-35033 Rennes, France 4- Institut Pasteur, CNRS – UMR_3738, Paris, France 5- CHU Rennes, Service de Génétique Clinique, CLAD Ouest, F-35033 Rennes, France 6- Université Paris-Saclay, UVSQ-INRA-ENVA, UMR-BREED, Montigny le Bretonneux, 78180, France 7- Fédération de Génétique, Laboratoire de Biologie Médicale, CHI de Poissy-St Germain en Laye, Poissy, 78300, France 8- CHU Rennes, Service de Biologie de la Reproduction-CECOS, F-35033 Rennes, France 9- Department of Paediatrics, University of Melbourne, Melbourne, VIC Australia *These authors should be regarded as co-senior authors of this study Corresponding authors: Sylvie Jaillard, [email protected]; Elena Tucker, [email protected] © The Author(s) 2020.
    [Show full text]
  • SGOL2 Antibody (Center) Affinity Purified Rabbit Polyclonal Antibody (Pab) Catalog # Ap20106c
    10320 Camino Santa Fe, Suite G San Diego, CA 92121 Tel: 858.875.1900 Fax: 858.622.0609 SGOL2 Antibody (Center) Affinity Purified Rabbit Polyclonal Antibody (Pab) Catalog # AP20106c Specification SGOL2 Antibody (Center) - Product Information Application WB,E Primary Accession Q562F6 Other Accession NP_689737.3 Reactivity Human Host Rabbit Clonality Polyclonal Isotype Rabbit Ig Calculated MW 144739 Antigen Region 391-419 SGOL2 Antibody (Center) - Additional Information Gene ID 151246 Other Names Shugoshin-like 2, Shugoshin-2, Sgo2, Tripin, SGOL2 Antibody (Center) (Cat. #AP20106c) SGOL2 western blot analysis in U-937 cell line lysates (35ug/lane).This demonstrates the Target/Specificity SGOL2 antibody detected the SGOL2 protein This SGOL2 antibody is generated from (arrow). rabbits immunized with a KLH conjugated synthetic peptide between 391-419 amino acids from the Central region of human SGOL2 Antibody (Center) - Background SGOL2. Cooperates with PPP2CA to protect Dilution centromeric cohesin from separase-mediated WB~~1:1000 cleavage in oocytes specifically during meiosis I. Has a crucial role in protecting REC8 at Format centromeres from cleavage by separase. Purified polyclonal antibody supplied in PBS with 0.09% (W/V) sodium azide. This During meiosis, protects centromeric cohesion antibody is purified through a protein A complexes until metaphase II/anaphase II column, followed by peptide affinity transition, preventing premature release of purification. meiosis-specific REC8 cohesin complexes from anaphase I centromeres. Is thus essential for Storage an accurate gametogenesis. May act by Maintain refrigerated at 2-8°C for up to 2 targeting PPP2CA to centromeres, thus leading weeks. For long term storage store at -20°C to cohesin dephosphorylation (By similarity).
    [Show full text]
  • Genome-Wide Variability in Recombination Activity Is Associated with Meiotic Chromatin Organization
    Downloaded from genome.cshlp.org on October 10, 2021 - Published by Cold Spring Harbor Laboratory Press Genome-wide variability in recombination activity is associated with meiotic chromatin organization Xiaofan Jin1, Geoff Fudenberg1,2, and Katherine S. Pollard1,3,4,* 1Gladstone Institutes, San Francisco, USA 2Department of Quantitative and Computational Biology, University of Southern California, Los Angeles, USA 3University of California San Francisco, San Francisco, USA 4Chan-Zuckerberg Biohub, San Francisco, USA *Corresponding author: [email protected] Running title: Recombination is related to chromatin structure Keywords: meiotic recombination, chromatin organization, 3D genome folding, PRDM9 binding, double strand breaks, crossovers 1 Downloaded from genome.cshlp.org on October 10, 2021 - Published by Cold Spring Harbor Laboratory Press 1 Abstract 2 Recombination enables reciprocal exchange of genomic information between parental chromo- 3 somes and successful segregation of homologous chromosomes during meiosis. Errors in this process 4 lead to negative health outcomes, while variability in recombination rate affects genome evolution. 5 In mammals, most crossovers occur in hotspots defined by PRDM9 motifs, though PRDM9 binding 6 peaks are not all equally hot. We hypothesize that dynamic patterns of meiotic genome folding are 7 linked to recombination activity. We apply an integrative bioinformatics approach to analyze how 8 three-dimensional (3D) chromosomal organization during meiosis relates to rates of double-strand- 9 break (DSB) and crossover (CO) formation at PRDM9 binding peaks. We show that active, spatially 10 accessible genomic regions during meiotic prophase are associated with DSB-favored loci, which fur- 11 ther adopt a transient locally active configuration in early prophase.
    [Show full text]
  • Casein Kinase 1 Is Required for Efficient Removal of Rec8 During Meiosis I
    REPORT REPORT Cell Cycle 9:13, 2657-2662; July 1, 2010; © 2010 Landes Bioscience Casein kinase 1 is required for efficient removal of Rec8 during meiosis I Cornelia Rumpf,1,† Lubos Cipak,1,† Andrej Dudas,1,† Zsigmond Benko,1 Miroslava Pozgajova,1 Christian G. Riedel,1-3 Gustav Ammerer,1 Karl Mechtler2 and Juraj Gregan1,* 1Max F. Perutz Laboratories; University of Vienna; Vienna, Austria; 2Research Institute of Molecular Pathology; Vienna, Austria; 3Department of Genetics; Harvard Medical School; Boston, MA USA †These authors contributed equally to this work. Key words: meiosis, chromosome segregation, cohesin, casein kinase, fission yeast Segregation of chromosomes during meiosis depends on separase cleavage of Rec8, the meiosis-specific alpha-kleisin subunit of cohesin. We mapped Rec8 phosphorylation sites by mass spectrometry and show that, in fission yeast, Rec8 phosphorylation is required for proper chromosome disjunction during meiosis. We further show that the fission yeast casein kinase 1 (CK1) delta/epsilon isoforms Hhp1 and Hhp2 are required for full levels of Rec8 phosphorylation and for efficient removal of Rec8 at the onset of anaphase I. Our data are consistent with the model that Hhp1/Hhp2-dependent phosphorylation of Rec8 is required for separase-mediated cleavage of Rec8 during meiosis I. Introduction Results Sexual reproduction depends on the generation of haploid gam- Mapping Rec8 phosphorylation sites. We purified Rec8-HA3 etes from diploid cells in a process called meiosis. The reduction from S. pombe cells harvested around metaphase I (4 hours after of chromosome number during meiosis is achieved by two suc- induction of synchronous meiosis) and mapped the phosphoryla- cessive nuclear divisions after one round of DNA replication.
    [Show full text]