DOCSLIB.ORG

Molecular Interactions of Desmin and VCP in Myofibrillar Myopathies

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Molecular Interactions of Desmin and VCP in Myofibrillar Myopathies Molecular Genetics of Primary Microcephaly and Microcephalic Primordial Dwarfism in Consanguineous Families from Pakistan Inaugural-Dissertation zur Erlangung des Doktorgrades der Mathematisch-Naturwissenschaftlichen Fakultät der Universität zu Köln vorgelegt von Ilyas Ahmad aus Dir Lower, Khyber Pukhtoonkhwa (Pakistan) 2016 Berichterstatter: Prof. Dr. Peter Nürnberg Berichterstatter: Prof. Dr. Angelika A. Noegel Tag der mündlichen Prüfung: June 30, 2015 The present research work was carried out from August 2011 to December 2014 at Cologne Center for Genomics under the supervision of Prof. Dr. Peter Nürnberg and at Institute of Biochemistry I, Medical Faculty at University of Cologne, Germany under the supervision of Prof. Dr. Angelika Noegel. Die vorliegende Arbeit wurde in der Zeit von August 2011 bis Dezember 2014 unter Anleitung von Prof. Dr. Peter Nürnberg am Cologne Center for Genomics und Prof. Dr. Angelika A. Noegel am Institut für Biochemie I der Medizinischen Fakultät der Universität zu Köln ausgeführt. This thesis is dedicated to my beloved mother, father, brothers and sisters Acknowledgements First and foremost, I would like to express my gratitude and appreciation to Prof. Dr. Peter Nürnberg, who has been an excellent supervisor and has supported me in every way throughout these four years. His active and engaged supervision was preeminent to finalize my PhD thesis. Thank you for your guidance, immense patience and valuable suggestions to complete this journey. I have had a lovely time in the lab and feel lucky to have had him as my mentor. My sincere indebtedness also goes to my co-supervisor Prof. Dr. Angelika Anna Noegel who has been helpful and patient to me; without her involvement, this work will not have been possible. Her encouragement and kindness will always be remembered. She was a tireless proofreader at various stages of this thesis. Thank you again for your quick feedback during the writing and revision process. I am also obliged to Prof. Dr. Hartmut Arndt and Dr. Holger Thiele for agreeing to be part of my exam committee on a very short notice. I also extends my sincere regards to Prof. Shahid Mahmmod Baig, National Institute for Biotechnology and Genetic Engineering (NIBGE), Pakistan for providing me the opportunity to work on this collaborative project. I am also thankful for providing their assistance and logistical support during my sampling visits in Pakistan. Also thanks given to Dr. Muhmmad Sajid Hussain for supervising all the experiments at Biochemistry 1 and always helpful with ideas over the last four years. Thanks for teaching me how to use adobe photoshop and image analysis, and to edit this thesis. Next, I would like to thank all the participating patients and their family members in this study. I would also extend my acknowledgments to the members of Human Molecular Genetics lab, NIBGE, Pakistan for their help in DNA extraction and gathering clinical information. I am also obliged to all the informants who gave me valuable information about patients during my field work. I warmly thank to all the present and past members of the CCG for providing such a warm working atmosphere. In particular I would like to thank: Dr. Mohammad Reza Toliat for whole-genome genotyping; Dr. Bigirt Budde and Gudrun Nürnberg for homozygosity mapping and linkage analysis; Dr. Holger Thiele and other members of bioinformatics team for whole exome sequencing; Hamid Shabbir for help with qPCR; Nina Dalibor, Elisabeth Kirst and Gerti Meyer for all technical assistance. I am sincerely thankful to Dr. Wilfred Gunia for their quick response to provide IT support and Holger Trucks and Dennis Lal for helpful discussion regarding various genetic tools and concepts. I am also thankful to all my colleagues in Biochemistry 1, particularly to Berthold for making antibodies and cell culture; Maria for introduction to confocal microscopy; Ralf Muller for providing different clones; Dr. Vivek Peche for experimental advices and encouragement; Martina Munck for technical assistance; office mates for great fun especially on birthday parties; Ilknur for suggestions during compiling my thesis. I am also thankful to all lab fellows, Sandra, Emrah, Salim and Kathrin. Appreciation also goes to Nicole Riedel and Dörte Püsche who made easy all the administrative work. I offer my acknowledgements to my friends Dr. Muhammad Tariq and Leigh Kroeger who proofread my thesis and made invaluable comments. Thanks to all my friends especially Arian Shir, Farman Afridi, Umar Dawar, Muhammad Arif, Asad Korejo, Raja Ghanzfar Ali, Mummad Ilyas and Usman outside this work with whom I have enjoyed these four years. My Acknowledgments could never adequately express the debt owed to my parents for their encouragement, love and unwavering support, and for that I dedicate this thesis to you. I deeply thank my brothers Iftikhar Ahmad and Ishfaq Ahmad and my sisters for their love and affection. This work was made possible with the financial support by the Center for Molecular Medicine Cologne (CMMC), University of Cologne, Germany. Table of contents 1. Abstract ....................................................................................................................... 4 2. Zusammenfassung ................................................................................................ 5 3.Introduction ................................................................................................................ 7 3.1 Pakistani Population .................................................................................................... 7 3.2 Microcephaly ............................................................................................................... 8 3.2.1 Primary microcephaly ........................................................................................... 9 3.2.1.1 Molecular genetics of primary microcephaly ..................................................10 3.2.2 Microcephalic primordial dwarfism (MPD) ............................................................13 3.2.2.1 Plk4 (Polo- like kinase 4) ...............................................................................15 3.3 The centrosome ..........................................................................................................17 3.3.1 Centrosome duplication cycle ..............................................................................18 3.4 Centrosome and primary microcephaly proteins .........................................................19 3.5 Short commentary on cellular pathways in microcephalic primordial dwarfism (MPD) 20 3.6 Corticogenesis and primary microcephaly ..................................................................21 4. Aims and objectives of the thesis .................................................................24 5. Materials and Methods .......................................................................................25 5.1 Families studied ..........................................................................................................25 5.2 Pedigree construction and analysis ............................................................................25 5.3 Blood sampling ...........................................................................................................27 5.4 DNA extraction ...........................................................................................................27 5.5 Exclusion mapping using short tandem repeats (STRs) ..........................................29 5.6 SNP genotyping methods .......................................................................................30 5.6.1 Axiom™ Genome-Wide CEU 1 Array ...............................................................31 5.6.2 Illumina HumanCoreExome-12 v1.1 BeadChip ................................................31 5.6.3 CytoScan HD Array..............................................................................................31 5.7 SNP array mapping and linkage analysis ....................................................................32 5.8 Sequencing reaction ...................................................................................................32 5.8.1 Sanger sequencing ..............................................................................................32 5.8.2 Pyrosequencing ...................................................................................................33 5.8.3 Whole-exome sequencing .......................................................................................35 5.9 RNA extraction ...........................................................................................................36 5.10 Cell culture and cell lines ..........................................................................................36 5.10.1 Establishment of patient primary fibroblasts .......................................................37 5.10.2 Cultivation of human fibroblasts .........................................................................37 5.10.3 Fibroblast synchronization at G2 phase .............................................................37 5.10.4 PLK4 construct and transfection ........................................................................38 5.10.5 Determination of growth .....................................................................................38 5.11 Immunofluorescence and microscopy .......................................................................38 5.11.1 Solutions used in Immunocytochemistry
Load more

Recommended publications
  • The Ciliated Cell Transcriptome A
    THE CILIATED CELL TRANSCRIPTOME A DISSERTATION SUBMITTED TO THE DEPARTMENT OF BIOLOGICAL SCIENCES AND THE COMMITTEE ON GRADUATE STUDIES OF STANFORD UNIVERSITY IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY Ramona Hoh March 2010 ! © 2010 by Ramona Amy Hoh. All Rights Reserved. Re-distributed by Stanford University under license with the author. This work is licensed under a Creative Commons Attribution- Noncommercial 3.0 United States License. http://creativecommons.org/licenses/by-nc/3.0/us/ This dissertation is online at: http://purl.stanford.edu/sk794dv5857 ii I certify that I have read this dissertation and that, in my opinion, it is fully adequate in scope and quality as a dissertation for the degree of Doctor of Philosophy. Timothy Stearns, Primary Adviser I certify that I have read this dissertation and that, in my opinion, it is fully adequate in scope and quality as a dissertation for the degree of Doctor of Philosophy. Mark Krasnow I certify that I have read this dissertation and that, in my opinion, it is fully adequate in scope and quality as a dissertation for the degree of Doctor of Philosophy. Maxence Nachury I certify that I have read this dissertation and that, in my opinion, it is fully adequate in scope and quality as a dissertation for the degree of Doctor of Philosophy. William Nelson Approved for the Stanford University Committee on Graduate Studies. Patricia J. Gumport, Vice Provost Graduate Education This signature page was generated electronically upon submission of this dissertation in electronic format. An original signed hard copy of the signature page is on file in University Archives.
    [Show full text]
  • Assembly of Centrosomal Proteins and Microtubule Organization Depends on PCM-1
    JCBArticle Assembly of centrosomal proteins and microtubule organization depends on PCM-1 Alexander Dammermann and Andreas Merdes Wellcome Trust Centre for Cell Biology, Institute of Cell and Molecular Biology, University of Edinburgh, Edinburgh EH9 3JR, UK he protein PCM-1 localizes to cytoplasmic granules treatment of cells with the microtubule inhibitor nocodazole. known as “centriolar satellites” that are partly enriched Inhibition or depletion of PCM-1 function further disrupted T around the centrosome. We inhibited PCM-1 function the radial organization of microtubules without affecting using a variety of approaches: microinjection of antibodies microtubule nucleation. Loss of microtubule organization into cultured cells, overexpression of a PCM-1 deletion was also observed after centrin or ninein depletion. Our mutant, and specific depletion of PCM-1 by siRNA. All data suggest that PCM-1–containing centriolar satellites are Downloaded from approaches led to reduced targeting of centrin, pericentrin, involved in the microtubule- and dynactin-dependent recruit- and ninein to the centrosome. Similar effects were seen ment of proteins to the centrosome, of which centrin and upon inhibition of dynactin by dynamitin, and after prolonged ninein are required for interphase microtubule organization. jcb.rupress.org Introduction Microtubule organization is essential for directional intra- the centrosomal surface or subsequently released and anchored cellular transport, for the modulation of cell morphology in other places of the cell (Mogensen, 1999). The initial step and locomotion, and for the formation of the spindle apparatus of microtubule nucleation is dependent on the function of during cell division. With the exception of plants, most cells 25S ring complexes of the protein ␥-tubulin and associated on December 31, 2017 organize their microtubule network using specialized structures, proteins (Zheng et al., 1995).
    [Show full text]
  • Specialized Cilia in Mammalian Sensory Systems
    Cells 2015, 4, 500-519; doi:10.3390/cells4030500 OPEN ACCESS cells ISSN 2073-4409 www.mdpi.com/journal/cells Review Specialized Cilia in Mammalian Sensory Systems Nathalie Falk, Marlene Lösl, Nadja Schröder and Andreas Gießl * Department of Biology, Animal Physiology, University of Erlangen-Nuremberg, 91058 Erlangen, Germany; E-Mails: [email protected] (N.F.); [email protected] (M.L.); [email protected] (A.G.) * Author to whom correspondence should be addressed; E-Mail: [email protected]; Tel.: +49-9131-85-28055; Fax: +49-9131-85-28060. Academic Editors: Gang Dong and William Tsang Received: 18 May 2015 / Accepted: 9 September 2015 / Published: 11 September 2015 Abstract: Cilia and flagella are highly conserved and important microtubule-based organelles that project from the surface of eukaryotic cells and act as antennae to sense extracellular signals. Moreover, cilia have emerged as key players in numerous physiological, developmental, and sensory processes such as hearing, olfaction, and photoreception. Genetic defects in ciliary proteins responsible for cilia formation, maintenance, or function underlie a wide array of human diseases like deafness, anosmia, and retinal degeneration in sensory systems. Impairment of more than one sensory organ results in numerous syndromic ciliary disorders like the autosomal recessive genetic diseases Bardet-Biedl and Usher syndrome. Here we describe the structure and distinct functional roles of cilia in sensory organs like the inner ear, the olfactory epithelium, and the retina of the mouse. The spectrum of ciliary function in fundamental cellular processes highlights the importance of elucidating ciliopathy-related proteins in order to find novel potential therapies.
    [Show full text]
  • Centrosome Cohesion: Functions of C-NAP1
    Provided by the author(s) and NUI Galway in accordance with publisher policies. Please cite the published version when available. Title Centrosome cohesion: functions of C-NAP1 Author(s) Flanagan, Anne-Marie Publication Date 2016-01-07 Item record http://hdl.handle.net/10379/5458 Downloaded 2021-09-25T13:41:15Z Some rights reserved. For more information, please see the item record link above. Centrosome Cohesion: Functions of C-NAP1 Anne-Marie Flanagan Centre for Chromosome Biology, School of Natural Sciences, National University of Ireland, Galway A thesis submitted to the National University of Ireland, Galway for the degree of Doctor of Philosophy September 2015 Supervisor: Prof. Ciaran Morrison Table of Contents Table of Contents ....................................................................................ii List of figures ........................................................................................... v List of tables ......................................................................................... viii Abreviations ............................................................................................ ix Acknowledgements ................................................................................xii Abstract ................................................................................................ xiii 1. Introduction .................................................................................... 14 1.1 Cell cycle overview ....................................................................................
    [Show full text]
  • Regulation of Centriolar Satellite Integrity and Its Physiology
    Cell. Mol. Life Sci. DOI 10.1007/s00018-016-2315-x Cellular and Molecular Life Sciences REVIEW Regulation of centriolar satellite integrity and its physiology 1,2 1,3 Akiko Hori • Takashi Toda Received: 19 May 2016 / Revised: 14 July 2016 / Accepted: 21 July 2016 Ó The Author(s) 2016. This article is published with open access at Springerlink.com Abstract Centriolar satellites comprise cytoplasmic gran- Keywords Cellular stress Á Centriole Á Ciliogenesis Á ules that are located around the centrosome. Their MSD1/SSX2IP Á Microtubule Á PCM1 Á PLK4 Á molecular identification was first reported more than a Phosphorylation Á Ubiquitylation quarter of a century ago. These particles are not static in the cell but instead constantly move around the centrosome. Abbreviations Over the last decade, significant advances in their molec- CS Centriolar satellites ular compositions and biological functions have been c-TuC c-Tubulin complex achieved due to comprehensive proteomics and genomics, HU Hydroxyurea super-resolution microscopy analyses and elegant genetic IR Ionising radiation manipulations. Centriolar satellites play pivotal roles in MCPH Microcephaly centrosome assembly and primary cilium formation MT Microtubule through the delivery of centriolar/centrosomal components MTOC Microtubule-organising centre from the cytoplasm to the centrosome. Their importance is PCM Pericentriolar material further underscored by the fact that mutations in genes SPB Spindle pole body encoding satellite components and regulators lead to vari- UV Ultraviolet ous human disorders such as ciliopathies. Moreover, the most recent findings highlight dynamic structural remod- elling in response to internal and external cues and unexpected positive feedback control that is exerted from Introduction the centrosome for centriolar satellite integrity.
    [Show full text]
  • Gene Ontology – Biological Process
    Table of Contents Abstract iv Acknowledgments vi Chapter 1: Introduction Long non-coding RNAs 1 Functions of long non-coding RNAs 4 Nuclear lncRNAs 4 Cytoplasmic lncRNAs 6 Long non-coding RNAs in cancer 9 Long non-coding RNAs in gliomas 12 Oncogenic long non-coding RNAs in gliomas 14 Tumor suppressor long non-coding RNAs in gliomas 17 References 19 Figures 33 Chapter 2: The functional role of APTR long non-coding RNA Abstract 35 Introduction 36 Results 38 Discussion 46 Materials and Methods 51 References 56 Figures 61 Chapter 3: Adapted from “LINC00152 promotes invasion in glioblastomas through a hairpin structure located near its 3’ end” My contribution to the paper 80 Abstract 82 Introduction 83 Results 84 ii Discussion 94 Materials and Methods 100 References 103 Figures 113 Chapter 4: Closing Remarks and Future Directions Alternative methods for discovering APTR interacting partners 136 APTR processing into different fragments 138 APTR and LINC00152 secondary structure 139 Identifying LINC00152 protein interactor 140 APTR as a regulator of gene expression 142 Molecular functions of LINC00152 144 Clinical potentials of APTR and LINC00152 145 References 146 Figures 152 iii Abstract Since the advent of high-throughput sequencing methods, long non-coding RNAs (lncRNAs) have been emerging as important players in physiological and pathological conditions. Among them, APTR (Alu-mediated p21 transcriptional regulator) is a lncRNA upregulated in tumors. This RNA regulates the cell cycle by recruiting the PRC2 protein complex (Polycomb Repressive Complex 2) to the promoter region of p21 gene, inducing its suppression. In addition, APTR and p21 expressions are anti-correlated in glioblastoma patient samples.
    [Show full text]
  • CDK5RAP2 Functions in Centrosome to Spindle Pole Attachment and DNA Damage Response
    JCB: Article CDK5RAP2 functions in centrosome to spindle pole attachment and DNA damage response Alexis R. Barr,1,2 John V. Kilmartin,3 and Fanni Gergely1,2 1Cancer Research UK Cambridge Research Institute, Cambridge CB2 0RE, England, UK 2Department of Oncology, University of Cambridge, Cambridge CB2 3RA, England, UK 3Medical Research Council Laboratory of Molecular Biology, Cambridge CB2 0QH, England, UK he centrosomal protein, CDK5RAP2, is mutated fail to recruit specific PCM components that mediate in primary microcephaly, a neurodevelopmental attachment to spindle poles. Furthermore, we show that Tdisorder characterized by reduced brain size. The the CNN1 domain enforces cohesion between parental Drosophila melanogaster homologue of CDK5RAP2, centrioles during interphase and promotes efficient DNA centrosomin (Cnn), maintains the pericentriolar matrix damage–induced G2 cell cycle arrest. Because mitotic (PCM) around centrioles during mitosis. In this study, we spindle positioning, asymmetric centrosome inheritance, demonstrate a similar role for CDK5RAP2 in vertebrate and DNA damage signaling have all been implicated cells. By disrupting two evolutionarily conserved domains in cell fate determination during neurogenesis, our find- of CDK5RAP2, CNN1 and CNN2, in the avian B cell line ings provide novel insight into how impaired CDK5RAP2 DT40, we find that both domains are essential for link- function could cause premature depletion of neural stem ing centrosomes to mitotic spindle poles. Although struc- cells and thereby microcephaly. turally intact, centrosomes lacking the CNN1 domain Introduction The centrosome consists of a pair of centrioles surrounded by 1995; Merdes et al., 1996, 2000; Gordon et al., 2001; Quintyne the pericentriolar matrix (PCM). In most animal cells, it is the and Schroer, 2002; Silk et al., 2009).
    [Show full text]
  • Genome-Wide Association Study to Find Modifiers for Tetralogy of Fallot in the 22Q11.2 Deletion Syndrome Identifies Variants in the GPR98 Locus on 5Q14.3
    1 2–4 * ) in an intron of −8 tetralogy of Fallot tetralogy of ◼ =2.98×10 P is the only gene that is known is the only gene that is known License, which permits use, distribution, License, which permits use, distribution, DOI: 10.1161/CIRCGENETICS.116.001690 MEF2C . Locus on 5q14.3 Data Supplement ivelo-cardio-facial syndrome ivelo-cardio-facial ◼ velocardiofacial syndrome/DiGeorge syndrome; Mendelian syndrome/DiGeorge velocardiofacial Inheritance in Man No. 192430, 188400) is one of the most births. common genomic disorders, occurring in 1 of 4000 live Over 90% of affected individuals have a de novo, hemizygous a de novo, have individuals 90% of affected Over 3 million base pair (Mb) deletion on chromosome 22q11.2. features of the syndrome, All subjects with the deletion have GPR98 1 genotype ◼ is published on behalf of the American Heart Association, Inc., by Wolters Kluwer Health, Wolters Association, Inc., by American Heart is published on behalf of the . 2017;10:e. DOI: 10.1161/CIRCGENETICS.116.001690.) Original Article (Myocyte-specific enhancer factor 2C). enhancer (Myocyte-specific Creative Commons Attribution Non-Commercial-NoDerivs Attribution Commons Creative MEF2C http://circgenetics.ahajournals.org/lookup/suppl/doi:10.1161/CIRCGENETICS.116.001690/-/DC1. DiGeorge syndrome DiGeorge Beverly S. Emanuel, PhD; Bernice E. Morrow, PhD; S. Emanuel, PhD; Bernice E. Morrow, Beverly ◼ , including (G-protein–coupled receptor V1) gene on chromosome 5q14.3. There was also suggestive evidence evidence also suggestive There was V1) gene on chromosome 5q14.3. (G-protein–coupled receptor Circ Cardiovasc Genet Circ Cardiovasc ( —To identify common genetic variants associated with TOF in individuals with 22q11.2DS, we in individuals TOF associated with identify common genetic variants —To is available at http://circgenetics.ahajournals.org is available Circulation: Cardiovascular Genetics Cardiovascular Circulation: GPR98 GPR98 chromosomes See Clinical Perspective Jonathan H.
    [Show full text]
  • Plk4-Induced Centriole Biogenesis in Human Cells
    Plk4-induced Centriole Biogenesis in Human Cells Dissertation zur Erlangung des Doktorgrades der Naturwissenschaften der Fakultät für Biologie der Ludwig-Maximilians Universität München Vorgelegt von Julia Kleylein-Sohn München, 2007 Dissertation eingereicht am: 27.11.2007 Tag der mündlichen Prüfung: 18.04.2008 Erstgutachter: Prof. E. A. Nigg Zweitgutachter: PD Dr. Angelika Böttger 2 Hiermit erkläre ich, dass ich die vorliegende Dissertation selbständig und ohne unerlaubte Hilfe angefertigt habe. Sämtliche Experimente wurden von mir selbst durchgeführt, soweit nicht explizit auf Dritte verwiesen wird. Ich habe weder an anderer Stelle versucht, eine Dissertation oder Teile einer solchen einzureichen bzw. einer Prüfungskommission vorzulegen, noch eine Doktorprüfung zu absolvieren. München, den 22.11.2007 3 TABLE OF CONTENTS SUMMARY…………………………………………………………………………..………. 6 INTRODUCTION……………………………………………………………………………. 7 Structure of the centrosome…………………………………………………………….. 7 The centrosome cycle…………………………………………………………………..10 Kinases involved in the regulation of centriole duplication………………………….12 Maintenance of centrosome numbers………………………………………………...13 Licensing of centriole duplication……………………………………………………... 15 ‘De novo ’ centriole assembly pathways in mammalian cells…………………..…...15 Templated centriole biogenesis in mammalian cells……………………………….. 18 The role of centrins and Sfi1p in centrosome duplication ……………………...…..19 Centriole biogenesis in C. elegans …………………………………………………… 21 Centriole biogenesis in human cells………………………………………………….. 23 Centrosome
    [Show full text]
  • Induction of Therapeutic Tissue Tolerance Foxp3 Expression Is
    Downloaded from http://www.jimmunol.org/ by guest on October 2, 2021 is online at: average * The Journal of Immunology , 13 of which you can access for free at: 2012; 189:3947-3956; Prepublished online 17 from submission to initial decision 4 weeks from acceptance to publication September 2012; doi: 10.4049/jimmunol.1200449 http://www.jimmunol.org/content/189/8/3947 Foxp3 Expression Is Required for the Induction of Therapeutic Tissue Tolerance Frederico S. Regateiro, Ye Chen, Adrian R. Kendal, Robert Hilbrands, Elizabeth Adams, Stephen P. Cobbold, Jianbo Ma, Kristian G. Andersen, Alexander G. Betz, Mindy Zhang, Shruti Madhiwalla, Bruce Roberts, Herman Waldmann, Kathleen F. Nolan and Duncan Howie J Immunol cites 35 articles Submit online. Every submission reviewed by practicing scientists ? is published twice each month by Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts http://jimmunol.org/subscription http://www.jimmunol.org/content/suppl/2012/09/17/jimmunol.120044 9.DC1 This article http://www.jimmunol.org/content/189/8/3947.full#ref-list-1 Information about subscribing to The JI No Triage! Fast Publication! Rapid Reviews! 30 days* Why • • • Material References Permissions Email Alerts Subscription Supplementary The Journal of Immunology The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2012 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. This information is current as of October 2, 2021.
    [Show full text]
  • Novel Approaches for Molecular Diagnosis of Genetic Diseases by Next Generation Sequencing
    Novel Approaches for Molecular Diagnosis of Genetic Diseases by Next Generation Sequencing: Application to Breast Cancer and Retinitis Pigmentosa in the Clinical Practice Work performed at the Molecular Genetics Unit of the Terrassa Hospital (Barcelona, Spain), and partially funded by the Carlos III Health Institute, the Fund for Health Research (FIS) from the Ministry of Health and Consumption of Spain and the Fundació Joan Costa Roma, of the Consorci Sanitari de Terrassa. The cover is composed of three images. The background image is an aerial view of the Douro River, a river that connects Spain with Portugal. The second image corresponds to a DNA sequence alignment software representation and the third one, used as a background for the titles and writing, is an image of the PicoTiter Plate after a sequencing run. Cover design by Miguel Constantino Silva de Sousa Dias. English edited by Richard May. Novel Approaches for Molecular Diagnosis of Genetic Diseases by Next Generation Sequencing: Application to Breast Cancer and Retinitis Pigmentosa in the Clinical Practice A Thesis Presented to The Department of Biochemistry and Molecular Biology Universitat Autònoma de Barcelona In Fulfilment of the Requirements for the Degree Philosophiæ Doctor (PhD) in Biochemistry, Molecular Biology and Biomedicine Director: Dr. José Miguel Carballo Villarino Tutor: Dr. Jaume Farrés Vicén by Miguel Miranda de Sousa Dias December 2013 El Dr. José Miguel Carballo Villarino, cap de la Unitat de Genètica Molecular de l’Hospital de Terrassa, i el Dr. Jaume
    [Show full text]
  • Human CPAP and CP110 in Centriole Elongation and Ciliogenesis
    Human CPAP and CP110 in Centriole Elongation and Ciliogenesis Dissertation zur Erlangung des Doktorgrades der Naturwissenschaften der Fakultät für Biologie der Ludwig-Maximilians Universität München Vorgelegt von Thorsten I. Schmidt München, 2010 Dissertation eingereicht am: 11.05.2010 Tag der mündlichen Prüfung: 25.10.2010 Erstgutachter: Prof. Dr. Erich A. Nigg Zweitgutachter: Prof. Dr. Angelika Böttger Hiermit erkläre ich, dass ich die vorliegende Dissertation selbständig und ohne unerlaubte Hilfe angefertigt habe. Sämtliche Experimente wurden von mir selbst durchgeführt, soweit nicht explizit auf Dritte verwiesen wird. Ich habe weder an anderer Stelle versucht, eine Dissertation oder Teile einer solchen einzureichen bzw. einer Prüfungskommission vorzulegen, noch eine Doktorprüfung zu absolvieren. München, den 11.05.2010 TABLE OF CONTENTS TABLE OF CONTENTS 1. SUMMARY............................................................................................................................1 2. INTRODUCTION .................................................................................................................2 2.1 Function and Structure of the Centrosome.....................................................................2 2.1.1 The Centrosome as MTOC in Proliferating Cells .................................................2 2.1.2 The Centriole as Template for Cilia and Flagella .................................................3 2.1.3 Molecular Composition and Structure of the Centrosome....................................3 2.2
    [Show full text]