DOCSLIB.ORG

The Mouse HP1 Proteins Are Not Required for Cell Viability but Are Essential For

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
The Mouse HP1 Proteins Are Not Required for Cell Viability but Are Essential For bioRxiv preprint doi: https://doi.org/10.1101/441279; this version posted October 11, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity.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 4.0 International license. 1 The mouse HP1 proteins are not required for cell viability but are essential for 2 preventing liver tumorigenesis 3 4 5 Nehmé Saksouk1-4*, Shefqet Hajdari1-4*, Marine Pratlong5, Célia Barrachina5, Aliki Zavoriti1-4, 6 Amélie Sarrazin6, Nelly Pirot1-4, Jean-Yohan Noël1-4, Lakhdar Khellaf4, Eric Fabbrizio1-4, 7, 7 Florence M. Cammas1-4, 7 8 9 1 IRCM, Institut de Recherche en Cancérologie de Montpellier, Montpellier, F-34298, France. 10 2 INSERM, U1194, Montpellier, F-34298, France. 11 3 Université de Montpellier, Montpellier, F-34090, France. 12 4 Institut régional du Cancer de Montpellier, Montpellier, F-34298, France. 13 5 MGX, Biocampus Montpellier, CNRS, INSERM, Univ Montpellier, Montpellier, France 14 6 MRI, BioCampus Montpellier, CNRS, INSERM, Univ Montpellier, Montpellier, France 15 7 CNRS, Route de Mende, Montpellier, France 16 17 Corresponding author: Florence Cammas, [email protected] 18 19 * Contributed equally to the work 1 bioRxiv preprint doi: https://doi.org/10.1101/441279; this version posted October 11, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity.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 4.0 International license. 20 Abstract 21 Chromatin organization plays essential roles in cell identity and functions. Here, we 22 demonstrated by hepatocyte-specific inactivation of the genes encoding the three 23 Heterochromatin Protein 1 (HP1α, β and γ) in mice (HP1-TKO) that these proteins are 24 dispensable for hepatocyte activity and survival. Conversely, the chronic absence of these 25 proteins led to a drastic increased incidence of liver tumor development. Molecular 26 characterization of HP1-TKO hepatocytes revealed that HP1 proteins are required for 27 maintenance of histone marks associated with heterochromatin, and for the appropriate 28 expression of large number of genes involved in liver-specific functions as well as in genes 29 encoding for transcriptional repressors of the KRAB-ZFP family. Moreover, several specific 30 endogenous retrovirus families were upregulated in HP1-TKO hepatocytes, leading to the 31 deregulated expression of genes in their vicinity. Our findings indicate that HP1 proteins act 32 as guardians of liver homeostasis to prevent tumor development through the modulation of 33 multiple chromatin-associated events. 34 Keywords: chromatin; HP1; cancer; liver; transcriptional silencing; endogenous retrovirus 35 36 2 bioRxiv preprint doi: https://doi.org/10.1101/441279; this version posted October 11, 2018. 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 4.0 International license. 37 Introduction 38 Chromatin is a large structure that is made of DNA fibers associated with histone 39 proteins, and that is essential for the interpretation of genetic information in a cell-type and 40 tissue-specific manner (Jenuwein and Allis 2001). Its alterations can have devastating 41 consequences, as evidenced by the large number of diseases induced by mutations in 42 chromatin-associated proteins (Koschmann et al. 2017; Mirabella et al. 2016), as well as by 43 the dramatic changes in chromatin organization observed in cancer cells (Stein et al. 2000). 44 Although extensively studied in the past three decades, it is still largely unknown how 45 chromatin organization is regulated and involved in whole organism homeostasis. 46 Chromatin can be divided, according to its structural and functional features, in 47 euchromatin and heterochromatin. Euchromatin displays low compaction level, is highly 48 enriched in genes, and is transcriptionally competent. Conversely heterochromatin is highly 49 compacted, enriched in repetitive DNA sequences, and mostly silent. Although, 50 heterochromatin was considered for a long time as an inert compartment with the single 51 function of silencing parasite DNA sequences, it is now recognized as a dynamic and 52 functional compartment (Janssen et al. 2018a). Heterochromatin Protein 1 (HP1) proteins 53 were first isolated as major heterochromatin components in Drosophila (James and Elgin 54 1986), and they are highly conserved from yeast to mammals which express three isoforms 55 (HP1α, HP1β and HP1γ). These proteins are characterized by an N-terminal chromodomain 56 (CD), which is involved in the recognition of the heterochromatin-associated histone mark H3 57 lysine-9 di- or trimethylated (H3K9me2/3), and a C-terminal chromoshadow domain (CSD), 58 which constitutes a platform for interaction with many protein partners. These two domains 59 are separated by the hinge domain that is crucial for HP1 association with RNA and 60 recruitment to heterochromatin (Eissenberg and Elgin 2014; Lomberk et al. 2006). Thus, HP1 61 proteins are at the crossroads of the structural and functional organization of chromatin. 62 Accordingly, HP1 are important for heterochromatin silencing, chromosome segregation, 63 regulation of gene expression, DNA repair and DNA replication (Dinant and Luijsterburg 3 bioRxiv preprint doi: https://doi.org/10.1101/441279; this version posted October 11, 2018. 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 4.0 International license. 64 2009; Fanti and Pimpinelli 2008; Nishibuchi and Nakayama 2014). Moreover, HP1 proteins 65 are essential for embryonic development in several organisms, including Drosophila 66 (Eissenberg et al. 1992), C. elegans (Schott et al. 2006) and the mouse (our unpublished 67 data). HP1α is essential for the plasticity of T helper (Th2) lymphocytes (Allan et al. 2012), 68 HP1β for neuro-muscular junctions (Aucott et al. 2008) and HP1γ for spermatogenesis (Abe 69 et al. 2011; Brown et al. 2010). Several studies also suggested a correlation between HP1 70 expression level and cancer development and/or metastasis formation; however, HP1 role in 71 tumorigenesis remains to be clarified (Dialynas et al. 2008; Vad-Nielsen and Nielsen 2015). 72 To assess the functions of this family of proteins in homeostasis and to bypass the 73 embryonic lethality of their genetic ablation, we inactivated all HP1 encoding genes in mouse 74 hepatocytes (HP1-TKO mice). Liver chromatin organization has been well characterized in 75 several physio-pathological conditions (Janssen et al. 2018b). In addition, several known 76 HP1 partners, including the transcription cofactors TRIM24 and TRIM28, and the histone- 77 lysine N-methyltransferase SUV39H1, play key roles in hepatocytes. Therefore, hepatocytes 78 constitute an excellent model to investigate HP1 roles in chromatin functions ((Herquel et al. 79 2011; Fan et al. 2013; Bojkowska et al. 2012a; Khetchoumian et al. 2007; Hardy and Mann 80 2016)). Unexpectedly, we found that in HP1-TKO mice, hepatocytes can differentiate and 81 contribute to liver functions throughout life. Conversely, HP1 proteins are critical for 82 preventing liver tumorigenesis. Finally, molecular characterization of HP1-TKO mice revealed 83 that HP1 are involved in the regulation of heterochromatin organization and of the expression 84 of genes and Endogenous Retroviruses (ERV). Altogether, these data highlight a new 85 function of HP1 proteins as guardians of liver homeostasis through the modulation of various 86 chromatin-associated events. 87 88 4 bioRxiv preprint doi: https://doi.org/10.1101/441279; this version posted October 11, 2018. 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 4.0 International license. 89 RESULTS 90 91 HP1 proteins are not required for hepatocyte proliferation and survival 92 To unravel HP1 functions in homeostasis, the HP1β and HP1γ encoding-genes (Cbx1 and 93 Cbx3, respectively) were inactivated in the liver of HP1αKO mice (Allan et al. 2012) using the 94 Cre recombinase expressed under the control of the hepatocyte-specific albumin promoter 95 (Postic et al. 1999) (Fig. 1A). These animals were called HP1-TKO. Liver-specific excision of 96 the Cbx1 and Cbx3 alleles was confirmed by PCR (Fig. 1B), and the absence of HP1β and 97 HP1γ protein expression was checked by western blotting. At 7 weeks post-partum as well 98 as at middle-aged, the overall level of HP1β and HP1γ was decreased by about 60% in HP1- 99 TKO liver (Fig. 1C). Furthermore, immuno-fluorescence analysis of liver cryosections using 100 anti-HP1β and -HP1γ antibodies showed that their expression was lost in about 60% of liver 101 cells in HP1-TKO mice compared with controls (wild type littermates or age-matched wild 102 type animals) (Fig. 1D). As this percentage corresponds to the hepatocyte fraction within the 103 liver, these findings indicate that both proteins were concomitantly depleted in most 104 hepatocytes. As expected since Cbx5 (the HP1α-encoding gene) was knocked out in all body 105 cells HP1α could not be detected HP1-TKO liver (Fig. 1B-C). Histological analysis of paraffin- 106 embedded liver tissue sections of 7-week-old (young) and 3-6-month-old (middle-aged) 107 control and HP1-TKO animals after hematoxylin/eosin/Safran staining did not reveal any 108 significant difference in the structural organization/morphology of hepatocytes and of the 109 parenchyma (Supplementary Figure 1). In agreement, analysis by immunohistochemistry 110 (IHC) of Tissue Micro Arrays (TMA) made of liver samples using anti-Ki67 and anti-activated 111 caspase 3 antibodies did not reveal any significant difference in cell proliferation and 112 apoptosis in young and middle-aged HP1-TKO and control mice (Fig.
Load more

Recommended publications
  • Maintenance of Self-Renewal Ability of Mouse Embryonic Stem Cells in The
    MaintenanceBlackwellMalden,GTCGenes1365-2443©?117OriginalDnmt1/3a/3bA 2006 TsumuraBlackwell to USA ArticleCells Publishing et Publishing al. triple knockoutInc Ltd ES cells of self-renewal ability of mouse embryonic stem cells in the absence of DNA methyltransferases Dnmt1, Dnmt3a and Dnmt3b Akiko Tsumura1,4, Tomohiro Hayakawa2, Yuichi Kumaki3,6, Shin-ichiro Takebayashi1, Morito Sakaue1, Chisa Matsuoka1, Kunitada Shimotohno4, Fuyuki Ishikawa5, En Li7, Hiroki R. Ueda3, Jun-ichi Nakayama2 and Masaki Okano1,* 1Laboratory for Mammalian Epigenetic Studies, 2Laboratory for Chromatin Dynamics, and 3Laboratory for Systems Biology, Center for Developmental Biology, RIKEN, 2-2-3 Minatojima-Minamimachi, Chuo-ku, Kobe 650-0047, Japan 4Department of Viral Oncology, Institute for Virus Research, Kyoto University, 53 Kawaharacho, Shogoin, Sakyo-ku, Kyoto 606-8501, Japan 5Department of Gene Mechanisms, Graduate School of Biostudies, Kyoto University, Kitashirakawa-Oiwake-cho, Sakyo-ku, Kyoto 606-8502, Japan 6INTEC Web and Genome Informatics Corp., 1-3-3 Shinsuna, Koto-ku, Tokyo 136-8637, Japan 7Epigenetics Program, Novartis Institute for Biomedical Research, 250 Massachusetts Ave., Cambridge, MA 02139, USA DNA methyltransferases Dnmt1, Dnmt3a and Dnmt3b cooperatively regulate cytosine methylation in CpG dinucleotides in mammalian genomes, providing an epigenetic basis for gene silencing and maintenance of genome integrity. Proper CpG methylation is required for the normal growth of various somatic cell types, indicating its essential role in the basic cellular function of mammalian cells. Previous studies using Dnmt1–/– or Dnmt3a–/–Dnmt3b–/– ES cells, however, have shown that undifferentiated embryonic stem (ES) cells can tolerate hypomethylation for their proliferation. In an attempt to investigate the effects of the complete loss of CpG DNA methyltransferase function, we established mouse ES cells lacking all three of these enzymes by gene targeting.
    [Show full text]
  • The Roles of Histone Deacetylase 5 and the Histone Methyltransferase Adaptor WDR5 in Myc Oncogenesis
    The Roles of Histone Deacetylase 5 and the Histone Methyltransferase Adaptor WDR5 in Myc oncogenesis By Yuting Sun This thesis is submitted in fulfilment of the requirements for the degree of Doctor of Philosophy at the University of New South Wales Children’s Cancer Institute Australia for Medical Research School of Women’s and Children’s Health, Faculty of Medicine University of New South Wales Australia August 2014 PLEASE TYPE THE UNIVERSITY OF NEW SOUTH WALES Thesis/Dissertation Sheet Surname or Family name: Sun First name: Yuting Other name/s: Abbreviation for degree as given in the University calendar: PhD School : School of·Women's and Children's Health Faculty: Faculty of Medicine Title: The Roles of Histone Deacetylase 5 and the Histone Methyltransferase Adaptor WDR5 in Myc oncogenesis. Abstract 350 words maximum: (PLEASE TYPE) N-Myc Induces neuroblastoma by regulating the expression of target genes and proteins, and N-Myc protein is degraded by Fbxw7 and NEDD4 and stabilized by Aurora A. The class lla histone deacetylase HDAC5 suppresses gene transcription, and blocks myoblast and leukaemia cell differentiation. While histone H3 lysine 4 (H3K4) trimethylation at target gene promoters is a pre-requisite for Myc· induced transcriptional activation, WDRS, as a histone H3K4 methyltransferase presenter, is required for H3K4 methylation and transcriptional activation mediated by a histone H3K4 methyltransferase complex. Here, I investigated the roles of HDAC5 and WDR5 in N-Myc overexpressing neuroblastoma. I have found that N-Myc upregulates HDAC5 protein expression, and that HDAC5 represses NEDD4 gene expression, increases Aurora A gene expression and consequently upregulates N-Myc protein expression in neuroblastoma cells.
    [Show full text]
  • (RRP1B) Modulates Metastasis Through Regulation of Histone Methylation
    Published OnlineFirst August 4, 2014; DOI: 10.1158/1541-7786.MCR-14-0167 Molecular Cancer Oncogenes and Tumor Suppressors Research Metastasis-Associated Protein Ribosomal RNA Processing 1 Homolog B (RRP1B) Modulates Metastasis through Regulation of Histone Methylation Minnkyong Lee1, Amy M. Dworkin1, Jens Lichtenberg1, Shashank J. Patel1, Niraj S. Trivedi2, Derek Gildea2, David M. Bodine1, and Nigel P.S. Crawford1 Abstract Overexpression of ribosomal RNA processing 1 homolog B (RRP1B) induces a transcriptional profile that accurately predicts patient outcome in breast cancer. However, the mechanism by which RRP1B modulates transcription is unclear. Here, the chromatin-binding properties of RRP1B were examined to define how it regulates metastasis-associated transcription. To identify genome-wide RRP1B-binding sites, high-throughput ChIP-seq was performed in the human breast cancer cell line MDA-MB-231 and HeLa cells using antibodies against endogenous RRP1B. Global changes in repressive marks such as histone H3 lysine 9 trimethylation (H3K9me3) were also examined by ChIP-seq. Analysis of these samples identified 339 binding regions in MDA- MB-231 cells and 689 RRP1B-binding regions in HeLa cells. Among these, 136 regions were common to both cell lines. Gene expression analyses of these RRP1B-binding regions revealed that transcriptional repression is the primary result of RRP1B binding to chromatin. ChIP-reChIP assays demonstrated that RRP1B co-occupies loci with decreased gene expression with the heterochromatin-associated proteins, tripartite motif-containing protein 28 (TRIM28/KAP1), and heterochromatin protein 1-a (CBX5/HP1a). RRP1B occupancy at these loci was also associated with higher H3K9me3 levels, indicative of heterochromatinization mediated by the TRIM28/HP1a complex.
    [Show full text]
  • Specificity, Propagation, and Memory of Pericentric Heterochromatin
    Article Specificity, propagation, and memory of pericentric heterochromatin Katharina Müller-Ott1, Fabian Erdel1, Anna Matveeva2, Jan-Philipp Mallm1, Anne Rademacher1, Matthias Hahn3, Caroline Bauer1, Qin Zhang2, Sabine Kaltofen1,†, Gunnar Schotta3, Thomas Höfer2 & Karsten Rippe1,* Abstract (Berger et al, 2009). These chromatin signals in turn recruit archi- tectural chromatin components or chromatin remodeling factors in The cell establishes heritable patterns of active and silenced chro- a highly dynamic manner and regulate genome access (McBryant matin via interacting factors that set, remove, and read epigenetic et al, 2006; Taverna et al, 2007; Campos & Reinberg, 2009; Clapier marks. To understand how the underlying networks operate, we & Cairns, 2009; Erdel et al, 2011a). On a global scale, the concerted have dissected transcriptional silencing in pericentric heterochro- and targeted activity of these networks results in the formation of matin (PCH) of mouse fibroblasts. We assembled a quantitative the denser, transcriptionally repressed heterochromatin state and map for the abundance and interactions of 16 factors related to the more open and biologically active euchromatin, which can be PCH in living cells and found that stably bound complexes of the distinguished at the resolution of the light microscope (Grewal & histone methyltransferase SUV39H1/2 demarcate the PCH state. Jia, 2007; Eissenberg & Reuter, 2009). A prototypic example for a From the experimental data, we developed a predictive mathe- constitutive heterochromatin domain is pericentric heterochromatin matical model that explains how chromatin-bound SUV39H1/2 (PCH) in mouse cells (Probst & Almouzni, 2008). It is characterized complexes act as nucleation sites and propagate a spatially by its high content of repetitive major satellite repeats and repres- confined PCH domain with elevated histone H3 lysine 9 trimethyla- sive epigenetic marks such as 5-methylcytosine (5meC) the binding tion levels via chromatin dynamics.
    [Show full text]
  • Genome-Wide DNA Methylation Analysis Reveals Molecular Subtypes of Pancreatic Cancer
    www.impactjournals.com/oncotarget/ Oncotarget, 2017, Vol. 8, (No. 17), pp: 28990-29012 Research Paper Genome-wide DNA methylation analysis reveals molecular subtypes of pancreatic cancer Nitish Kumar Mishra1 and Chittibabu Guda1,2,3,4 1Department of Genetics, Cell Biology and Anatomy, University of Nebraska Medical Center, Omaha, NE, 68198, USA 2Bioinformatics and Systems Biology Core, University of Nebraska Medical Center, Omaha, NE, 68198, USA 3Department of Biochemistry and Molecular Biology, University of Nebraska Medical Center, Omaha, NE, 68198, USA 4Fred and Pamela Buffet Cancer Center, University of Nebraska Medical Center, Omaha, NE, 68198, USA Correspondence to: Chittibabu Guda, email: [email protected] Keywords: TCGA, pancreatic cancer, differential methylation, integrative analysis, molecular subtypes Received: October 20, 2016 Accepted: February 12, 2017 Published: March 07, 2017 Copyright: Mishra et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC-BY), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. ABSTRACT Pancreatic cancer (PC) is the fourth leading cause of cancer deaths in the United States with a five-year patient survival rate of only 6%. Early detection and treatment of this disease is hampered due to lack of reliable diagnostic and prognostic markers. Recent studies have shown that dynamic changes in the global DNA methylation and gene expression patterns play key roles in the PC development; hence, provide valuable insights for better understanding the initiation and progression of PC. In the current study, we used DNA methylation, gene expression, copy number, mutational and clinical data from pancreatic patients.
    [Show full text]
  • Lamina-Associated Domains: Peripheral Matters and Internal Affairs Nolwenn Briand1,2 and Philippe Collas1,2*
    Briand and Collas Genome Biology (2020) 21:85 https://doi.org/10.1186/s13059-020-02003-5 REVIEW Open Access Lamina-associated domains: peripheral matters and internal affairs Nolwenn Briand1,2 and Philippe Collas1,2* * Correspondence: philippe.collas@ medisin.uio.no Abstract 1Department of Molecular Medicine, Faculty of Medicine, University of At the nuclear periphery, associations of chromatin with the nuclear lamina through Oslo, PO Box 1112 Blindern, 0317 lamina-associated domains (LADs) aid functional organization of the genome. We Oslo, Norway review the organization of LADs and provide evidence of LAD heterogeneity from 2Department of Immunology and Transfusion Medicine, Oslo cell ensemble and single-cell data. LADs are typically repressive environments in the University Hospital, 0424 Oslo, genome; nonetheless, we discuss findings of lamin interactions with regulatory Norway elements of active genes, and the role lamins may play in genome regulation. We address the relationship between LADs and other genome organizers, and the involvement of LADs in laminopathies. The current data lay the basis for future studies on the significance of lamin-chromatin interactions in health and disease. Keywords: 3D genome, Chromatin, LAD, Lamin A mutation, Nuclear envelope, Nuclear lamin, Radial positioning Introduction A snapshot of the genome in three dimensions In the eukaryotic nucleus, individual chromosomes occupy non-random territories and a radial (i.e., center-to-periphery) disposition enabled by sequestering specific chromo- somal domains to the nuclear envelope. Current views of the three-dimensional (3D) genome depict a hierarchical architecture of chromatin [1, 2]. Chromosome conform- ation capture combined with high-throughput sequencing (a method called Hi-C [3]) makes it possible to map interactions within and between chromosomes across ge- nomes [4].
    [Show full text]
  • H19 Lncrna Controls Gene Expression of the Imprinted Gene Network by Recruiting MBD1
    H19 lncRNA controls gene expression of the Imprinted Gene Network by recruiting MBD1 Paul Monniera,b, Clémence Martineta, Julien Pontisc, Irina Stanchevad, Slimane Ait-Si-Alic, and Luisa Dandoloa,1 aGenetics and Development Department, Institut National de la Santé et de la Recherche Médicale U1016, Centre National de la Recherche Scientifique (CNRS) Unité Mixte de Recherche (UMR) 8104, University Paris Descartes, Institut Cochin, Paris 75014, France; bUniversity Paris Pierre and Marie Curie, Paris 75005, France; cUniversity Paris Diderot, Sorbonne Paris Cité, Laboratoire Epigénétique et Destin Cellulaire, CNRS UMR 7216, Paris 75013, France; and dWellcome Trust Centre for Cell Biology, University of Edinburgh, Edinburgh EH9 3JR, United Kingdom Edited by Marisa Bartolomei, University of Pennsylvania, Philadelphia, PA, and accepted by the Editorial Board November 11, 2013 (received for review May 30, 2013) The H19 gene controls the expression of several genes within the this control is transcriptional or posttranscriptional and whether Imprinted Gene Network (IGN), involved in growth control of the these nine targets are direct or indirect targets remain elusive. embryo. However, the underlying mechanisms of this control re- Several lncRNAs interact with chromatin-modifying com- main elusive. Here, we identified the methyl-CpG–binding domain plexes and appear to exert a transcriptional control by targeting fi protein 1 MBD1 as a physical and functional partner of the H19 local chromatin modi cations at discrete genomic regions (8, 9). long noncoding RNA (lncRNA). The H19 lncRNA–MBD1 complex is In the case of imprinted clusters, the DMRs controlling the ex- fi pression of imprinted genes exhibit parent-of-origin epigenetic required for the control of ve genes of the IGN.
    [Show full text]
  • Prognostic Values of Distinct CBX Family Members in Breast Cancer Liang, Yuan-Ke; Lin, Hao-Yu; Chen, Chun-Fa; Zeng, De
    University of Groningen Prognostic values of distinct CBX family members in breast cancer Liang, Yuan-Ke; Lin, Hao-Yu; Chen, Chun-Fa; Zeng, De Published in: Oncotarget DOI: 10.18632/oncotarget.21325 IMPORTANT NOTE: You are advised to consult the publisher's version (publisher's PDF) if you wish to cite from it. Please check the document version below. Document Version Publisher's PDF, also known as Version of record Publication date: 2017 Link to publication in University of Groningen/UMCG research database Citation for published version (APA): Liang, Y-K., Lin, H-Y., Chen, C-F., & Zeng, D. (2017). Prognostic values of distinct CBX family members in breast cancer. Oncotarget, 8(54), 92375-92387. https://doi.org/10.18632/oncotarget.21325 Copyright Other than for strictly personal use, it is not permitted to download or to forward/distribute the text or part of it without the consent of the author(s) and/or copyright holder(s), unless the work is under an open content license (like Creative Commons). The publication may also be distributed here under the terms of Article 25fa of the Dutch Copyright Act, indicated by the “Taverne” license. More information can be found on the University of Groningen website: https://www.rug.nl/library/open-access/self-archiving-pure/taverne- amendment. Take-down policy If you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediately and investigate your claim. Downloaded from the University of Groningen/UMCG research database (Pure): http://www.rug.nl/research/portal.
    [Show full text]
  • MBD1 Antibody
    BioVision 09/14 For research use only MBD1 Antibody ChIP assays were performed using U2OS cells and the antibody and ALTERNATE NAMES: CXXC3, PCM1, RFT, Methyl-CpG-Binding Domain 1 optimized PCR primer sets for qPCR. Beads only were used as a negative CATALOG #: 6828-25 control. The Fig shows the recovery, expressed as a % of input (the relative AMOUNT: 25 µg amount of IP DNA compared to input DNA after qPCR analysis). HOST/ISOTYPE: Rabbit IMMUNOGEN: MBD1 (Methyl-CpG-binding domain protein 1) synthetic peptide containing a sequence from the N-terminal conjugated to KLH FORM: Liquid FORMULATION: In PBS containing 0.05% azide and 0.05% ProClin 300. PURIFICATION: Affinity purified SPECIES REACTIVITY: Human. STORAGE CONDITIONS: Store at -20°C; for long storage, store at -80°C. Avoid multiple freeze-thaw cycles. To determine the titer, an ELISA was performed using a serial dilution of the DESCRIPTION: MBD1 is a transcriptional repressor that specifically binds to methylated antibody. The antigen used was a CpG dinucleotides in promoter sequences. MBD1 acts by recruiting a variety of histone peptide containing the histone deacetylases (HDAC’s) and chromatin remodelling factors. MBD1-dependent transcriptional modification of interest. By plotting the repression is mediated by ATF7IP through the recruitment of factors such as the histone absorbance against the antibody methyltransferase SETDB1. MBD1 probably forms a complex with SETDB1 and ATF7IP dilution the titer of the antibody was which couples DNA methylation to H3K9 trimethylation and represses transcription. estimated to be 1:20,000. APPLICATION: ChIP: 1.5 µg/ChIP, WB: 1:500, ELISA: 1:1000.
    [Show full text]
  • Lamin B1 Is Required for Mature Neuron-Specific Gene Expression
    ARTICLE Received 4 Jul 2016 | Accepted 28 Feb 2017 | Published 20 Apr 2017 DOI: 10.1038/ncomms15098 OPEN Lamin B1 is required for mature neuron-specific gene expression during olfactory sensory neuron differentiation Crystal M. Gigante1,2, Michele Dibattista3,4, Frederick N. Dong1, Xiaobin Zheng2, Sibiao Yue2, Stephen G. Young5, Johannes Reisert3, Yixian Zheng2 & Haiqing Zhao1 B-type lamins are major constituents of the nuclear lamina in all metazoan cells, yet have specific roles in the development of certain cell types. Although they are speculated to regulate gene expression in developmental contexts, a direct link between B-type lamins and developmental gene expression in an in vivo system is currently lacking. Here, we identify lamin B1 as a key regulator of gene expression required for the formation of functional olfactory sensory neurons. By using targeted knockout in olfactory epithelial stem cells in adult mice, we show that lamin B1 deficient neurons exhibit attenuated response to odour stimulation. This deficit can be explained by decreased expression of genes involved in mature neuron function, along with increased expression of genes atypical of the olfactory lineage. These results support that the broadly expressed lamin B1 regulates expression of a subset of genes involved in the differentiation of a specific cell type. 1 Department of Biology, The Johns Hopkins University, Baltimore, Maryland 21218, USA. 2 Department of Embryology, Carnegie Institution for Science, Baltimore, Maryland 21218, USA. 3 Monell Chemical Senses Center, Philadelphia, Pennsylvania 19104, USA. 4 Department of Basic Medical Sciences, Neuroscience and Sensory Organs, University of Bari ‘A. Moro’, Bari 70121, Italy. 5 Department of Medicine, Molecular Biology Institute and Department of Human Genetics, University of California, Los Angeles, California 90095, USA.
    [Show full text]
  • The Genetic Program of Pancreatic Beta-Cell Replication in Vivo
    Page 1 of 65 Diabetes The genetic program of pancreatic beta-cell replication in vivo Agnes Klochendler1, Inbal Caspi2, Noa Corem1, Maya Moran3, Oriel Friedlich1, Sharona Elgavish4, Yuval Nevo4, Aharon Helman1, Benjamin Glaser5, Amir Eden3, Shalev Itzkovitz2, Yuval Dor1,* 1Department of Developmental Biology and Cancer Research, The Institute for Medical Research Israel-Canada, The Hebrew University-Hadassah Medical School, Jerusalem 91120, Israel 2Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot, Israel. 3Department of Cell and Developmental Biology, The Silberman Institute of Life Sciences, The Hebrew University of Jerusalem, Jerusalem 91904, Israel 4Info-CORE, Bioinformatics Unit of the I-CORE Computation Center, The Hebrew University and Hadassah, The Institute for Medical Research Israel- Canada, The Hebrew University-Hadassah Medical School, Jerusalem 91120, Israel 5Endocrinology and Metabolism Service, Department of Internal Medicine, Hadassah-Hebrew University Medical Center, Jerusalem 91120, Israel *Correspondence: [email protected] Running title: The genetic program of pancreatic β-cell replication 1 Diabetes Publish Ahead of Print, published online March 18, 2016 Diabetes Page 2 of 65 Abstract The molecular program underlying infrequent replication of pancreatic beta- cells remains largely inaccessible. Using transgenic mice expressing GFP in cycling cells we sorted live, replicating beta-cells and determined their transcriptome. Replicating beta-cells upregulate hundreds of proliferation- related genes, along with many novel putative cell cycle components. Strikingly, genes involved in beta-cell functions, namely glucose sensing and insulin secretion were repressed. Further studies using single molecule RNA in situ hybridization revealed that in fact, replicating beta-cells double the amount of RNA for most genes, but this upregulation excludes genes involved in beta-cell function.
    [Show full text]
  • Mice Lacking Methyl-Cpg Binding Protein 1 Have Deficits in Adult Neurogenesis and Hippocampal Function
    Mice lacking methyl-CpG binding protein 1 have deficits in adult neurogenesis and hippocampal function Xinyu Zhao*, Tetsuya Ueba*†, Brian R. Christie‡, Basam Barkho*, Michael J. McConnell§, Kinichi Nakashima*, Edward S. Lein*, Brennan D. Eadie‡, Andrew R. Willhoite*, Alysson R. Muotri*, Robert G. Summers*, Jerold Chun§, Kuo-Fen Lee¶, and Fred H. Gage*ʈ *Laboratory of Genetics and ¶Peptide Biology Laboratory, The Salk Institute for Biological Studies, La Jolla, CA 92037; §Department of Pharmacology, University of California, 9500 Gilman Drive, La Jolla, CA 92093; and ‡Department of Psychology, University of British Columbia, 2136 West Mall, Vancouver, BC, Canada V6T 1Z4 Communicated by Inder M. Verma, The Salk Institute for Biological Studies, San Diego, CA, April 2, 2003 (received for review March 7, 2003) DNA methylation-mediated epigenetic regulation plays critical roles in vivo function of MBD1 is still unclear. Because MBD1 has no in regulating mammalian gene expression, but its role in normal brain sequence specificity, except for methylated CpG, its in vivo target function is not clear. Methyl-CpG binding protein 1 (MBD1), a member genes are not known. of the methylated DNA-binding protein family, has been shown to Maintaining normal DNA methylation levels is critical during bind methylated gene promoters and facilitate transcriptional repres- mammalian development, as demonstrated by the embryonic Ϫ Ϫ sion in vitro. Here we report the generation and analysis of MBD1؊/؊ lethality of Dnmt1 / mice (17). Mutation of the de novo DNA -mice. MBD1؊/؊ mice had no detectable developmental defects and methyltransferase 1-36 (Dnmt3b) causes immunodeficiency, cen -appeared healthy throughout life. However, we found that MBD1؊/؊ tromeric instability, and facial anomalies (ICF) syndrome in hu neural stem cells exhibited reduced neuronal differentiation and mans, with characteristics of genomic instability (18).
    [Show full text]