DOCSLIB.ORG

The Ubiquitination Enzymes of Leishmania Mexicana

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
The Ubiquitination Enzymes of Leishmania Mexicana The ubiquitination enzymes of Leishmania mexicana Rebecca Jayne Burge Doctor of Philosophy University of York Biology October 2020 Abstract Post-translational modifications such as ubiquitination are important for orchestrating the cellular transformations that occur as the Leishmania parasite differentiates between its main morphological forms, the promastigote and amastigote. Although 20 deubiquitinating enzymes (DUBs) have been partially characterised in Leishmania mexicana, little is known about the role of E1 ubiquitin-activating (E1), E2 ubiquitin- conjugating (E2) and E3 ubiquitin ligase (E3) enzymes in this parasite. Using bioinformatic methods, 2 E1, 13 E2 and 79 E3 genes were identified in the L. mexicana genome. Subsequently, bar-seq analysis of 23 E1, E2 and HECT/RBR E3 null mutants generated in promastigotes using CRISPR-Cas9 revealed that the E2s UBC1/CDC34, UBC2 and UEV1 and the HECT E3 ligase HECT2 are required for successful promastigote to amastigote differentiation and UBA1b, UBC9, UBC14, HECT7 and HECT11 are required for normal proliferation during mouse infection. Null mutants could not be generated for the E1 UBA1a or the E2s UBC3, UBC7, UBC12 and UBC13, suggesting these genes are essential in promastigotes. X-ray crystal structure analysis of UBC2 and UEV1, orthologues of human UBE2N and UBE2V1/UBE2V2 respectively, revealed a heterodimer with a highly conserved structure and interface. Furthermore, recombinant L. mexicana UBA1a was found to load ubiquitin onto UBC2, allowing UBC2- UEV1 to form K63-linked di-ubiquitin chains in vitro. UBC2 was also shown to cooperate with human E3s RNF8 and BIRC2 in vitro to form non-K63-linked polyubiquitin chains, but association of UBC2 with UEV1 inhibits this ability. Using affinity purification proteomics, 3 putative UBC2:E3 pairs and potential substrates of UBC2 and UEV1, which include proteins associated with cellular respiration, intracellular transport and pH regulation, were identified. Therefore, the essential requirement for UBC2 and UEV1 in promastigote to amastigote differentiation could be explained by roles for UBC2-UEV1- mediated ubiquitination in regulating the changes in metabolism and protein trafficking that occur during this transition. 2 Table of contents Title page ...................................................................................................................... 1 Abstract ........................................................................................................................ 2 Table of contents .......................................................................................................... 3 List of tables ................................................................................................................. 6 List of figures ................................................................................................................ 7 List of accompanying material....................................................................................... 9 Acknowledgements ..................................................................................................... 10 Author’s declaration .................................................................................................... 12 1 Introduction .......................................................................................................... 13 1.1 Leishmania and leishmaniasis ...................................................................... 13 Epidemiology ......................................................................................... 13 Disease pathology ................................................................................. 13 Life cycle overview ................................................................................ 15 Cellular composition .............................................................................. 17 Gene expression ................................................................................... 19 Current treatments ................................................................................. 21 Genetic approaches to the study of Leishmania .................................... 23 1.2 Ubiquitination in non-trypanosomatids .......................................................... 24 Introduction to the ubiquitin system........................................................ 24 Diversity and roles of ubiquitin modifications ......................................... 26 E1 ubiquitin-activating enzymes ............................................................ 28 E2 ubiquitin-conjugating enzymes ......................................................... 29 E3 ubiquitin ligases ................................................................................ 31 Deubiquitinating enzymes ...................................................................... 35 The proteasome .................................................................................... 36 Ubiquitin-like modifiers ........................................................................... 37 1.3 Ubiquitination in trypanosomatids ................................................................. 37 Discovery of the trypanosomatid ubiquitin system ................................. 37 Ubiquitination in the life cycle of trypanosomatids .................................. 39 Roles for ubiquitin in trypanosomatid endocytosis ................................. 41 Ubiquitination and the infection process ................................................ 42 Trypanosomatid ubiquitin system inhibitors ........................................... 43 Ubls in trypanosomatids ........................................................................ 44 1.4 Aims ............................................................................................................. 46 2 Materials and methods ........................................................................................ 47 2.1 Bioinformatics ............................................................................................... 47 Protein domain searches ....................................................................... 47 Other searches ...................................................................................... 47 2.2 Molecular biology .......................................................................................... 48 Polymerase chain reaction (PCR) .......................................................... 48 Agarose gel electrophoresis .................................................................. 48 Transformations ..................................................................................... 49 Bacterial culture and storage ................................................................. 49 DNA extraction from E. coli .................................................................... 49 Restriction digestion .............................................................................. 49 DNA sequencing .................................................................................... 50 2.3 Leishmania cell culture ................................................................................. 50 Promastigote cell culture ....................................................................... 50 Promastigote to amastigote differentiation ............................................. 50 Cell counting .......................................................................................... 51 Axenic amastigote viability assay .......................................................... 51 3 Transfection and selection ..................................................................... 51 CRISPR-Cas9-based endogenous tagging ............................................ 52 Live cell imaging .................................................................................... 53 CRISPR-Cas9-based null mutant generation ......................................... 53 Genomic DNA extraction ....................................................................... 53 Pooled bar-seq screen ........................................................................... 53 2.4 Biochemical techniques ................................................................................ 55 Activity-based protein profiling ............................................................... 55 Generation of E. coli expression plasmids ............................................. 56 Bacterial expression .............................................................................. 56 Bacterial cell lysis .................................................................................. 57 Nickel affinity purification ....................................................................... 57 Tag cleavage and removal ..................................................................... 57 Size-exclusion chromatography ............................................................. 57 Assessment of protein purity and storage .............................................. 58 Size Exclusion Chromatography-Multi-Angle Laser Light Scattering (SEC- MALLS) 58 SDS-PAGE ............................................................................................ 58 Western blotting ..................................................................................... 59 Thioester assay ..................................................................................... 59 Diubiquitin formation assay ...................................................................
Load more

Recommended publications
  • Ubiquitin-Conjugating Enzyme UBE2J1 Negatively Modulates
    Feng et al. Virology Journal (2018) 15:132 https://doi.org/10.1186/s12985-018-1040-5 RESEARCH Open Access Ubiquitin-conjugating enzyme UBE2J1 negatively modulates interferon pathway and promotes RNA virus infection Tingting Feng1, Lei Deng1, Xiaochuan Lu1, Wen Pan1, Qihan Wu2* and Jianfeng Dai1,2* Abstract Background: Viral infection activates innate immune pathways and interferons (IFNs) play a pivotal role in the outcome of a viral infection. Ubiquitin modifications of host and viral proteins significantly influence the progress of virus infection. Ubiquitin-conjugating enzyme E2s (UBE2) have the capacity to determine ubiquitin chain topology and emerge as key mediators of chain assembly. Methods: In this study, we screened the functions of 34 E2 genes using an RNAi library during Dengue virus (DENV) infection. RNAi and gene overexpression approaches were used to study the gene function in viral infection and interferon signaling. Results: We found that silencing UBE2J1 significantly impaired DENV infection, while overexpression of UBE2J1 enhanced DENV infection. Further studies suggested that type I IFN expression was significantly increased in UBE2J1 silenced cells and decreased in UBE2J1 overexpressed cells. Reporter assay suggested that overexpression of UBE2J1 dramatically suppressed RIG-I directed IFNβ promoter activation. Finally, we have confirmed that UBE2J1 can facilitate the ubiquitination and degradation of transcription factor IFN regulatory factor 3 (IRF3). Conclusion: These results suggest that UBE2 family member UBE2J1 can negatively regulate type I IFN expression, thereby promote RNA virus infection. Keywords: UBE2J1, Dengue virus, Interferons, IRF3, K48 ubiquitination Background antiviral responses [3]. IFN-α/β regulates the synthesis Dengue virus (DENV), transmitted by Aedes aegypti and of antiviral proteins and immunoregulatory factors Aedes albopicuts, causes an emerging tropical disease through the JAK/STAT signaling pathway [4, 5].
    [Show full text]
  • Identification of the Binding Partners for Hspb2 and Cryab Reveals
    Brigham Young University BYU ScholarsArchive Theses and Dissertations 2013-12-12 Identification of the Binding arP tners for HspB2 and CryAB Reveals Myofibril and Mitochondrial Protein Interactions and Non- Redundant Roles for Small Heat Shock Proteins Kelsey Murphey Langston Brigham Young University - Provo Follow this and additional works at: https://scholarsarchive.byu.edu/etd Part of the Microbiology Commons BYU ScholarsArchive Citation Langston, Kelsey Murphey, "Identification of the Binding Partners for HspB2 and CryAB Reveals Myofibril and Mitochondrial Protein Interactions and Non-Redundant Roles for Small Heat Shock Proteins" (2013). Theses and Dissertations. 3822. https://scholarsarchive.byu.edu/etd/3822 This Thesis is brought to you for free and open access by BYU ScholarsArchive. It has been accepted for inclusion in Theses and Dissertations by an authorized administrator of BYU ScholarsArchive. For more information, please contact [email protected], [email protected]. Identification of the Binding Partners for HspB2 and CryAB Reveals Myofibril and Mitochondrial Protein Interactions and Non-Redundant Roles for Small Heat Shock Proteins Kelsey Langston A thesis submitted to the faculty of Brigham Young University in partial fulfillment of the requirements for the degree of Master of Science Julianne H. Grose, Chair William R. McCleary Brian Poole Department of Microbiology and Molecular Biology Brigham Young University December 2013 Copyright © 2013 Kelsey Langston All Rights Reserved ABSTRACT Identification of the Binding Partners for HspB2 and CryAB Reveals Myofibril and Mitochondrial Protein Interactors and Non-Redundant Roles for Small Heat Shock Proteins Kelsey Langston Department of Microbiology and Molecular Biology, BYU Master of Science Small Heat Shock Proteins (sHSP) are molecular chaperones that play protective roles in cell survival and have been shown to possess chaperone activity.
    [Show full text]
  • A Drosophila Ortholog of the Human Cylindromatosis Tumor Suppressor
    RESEARCH ARTICLE 2605 Development 134, 2605-2614 (2007) doi:10.1242/dev.02859 A Drosophila ortholog of the human cylindromatosis tumor suppressor gene regulates triglyceride content and antibacterial defense Theodore Tsichritzis1, Peer C. Gaentzsch3, Stylianos Kosmidis2, Anthony E. Brown3, Efthimios M. Skoulakis2, Petros Ligoxygakis3,* and George Mosialos1,4,* The cylindromatosis (CYLD) gene is mutated in human tumors of skin appendages. It encodes a deubiquitylating enzyme (CYLD) that is a negative regulator of the NF-␬B and JNK signaling pathways, in vitro. However, the tissue-specific function and regulation of CYLD in vivo are poorly understood. We established a genetically tractable animal model to initiate a systematic investigation of these issues by characterizing an ortholog of CYLD in Drosophila. Drosophila CYLD is broadly expressed during development and, in adult animals, is localized in the fat body, ovaries, testes, digestive tract and specific areas of the nervous system. We demonstrate that the protein product of Drosophila CYLD (CYLD), like its mammalian counterpart, is a deubiquitylating enzyme. Impairment of CYLD expression is associated with altered fat body morphology in adult flies, increased triglyceride levels and increased survival under starvation conditions. Furthermore, flies with compromised CYLD expression exhibited reduced resistance to bacterial infections. All mutant phenotypes described were reversible upon conditional expression of CYLD transgenes. Our results implicate CYLD in a broad range of functions associated with fat homeostasis and host defence in Drosophila. KEY WORDS: Cylindromatosis, Drosophila, Fat body, Host defense, NF-kappaB INTRODUCTION disease and it is required for the proper development of T Familial cylindromatosis is an autosomal-dominant predisposition lymphocytes in mice (Costello et al., 2005; Reiley et al., 2006).
    [Show full text]
  • A Computational Approach for Defining a Signature of Β-Cell Golgi Stress in Diabetes Mellitus
    Page 1 of 781 Diabetes A Computational Approach for Defining a Signature of β-Cell Golgi Stress in Diabetes Mellitus Robert N. Bone1,6,7, Olufunmilola Oyebamiji2, Sayali Talware2, Sharmila Selvaraj2, Preethi Krishnan3,6, Farooq Syed1,6,7, Huanmei Wu2, Carmella Evans-Molina 1,3,4,5,6,7,8* Departments of 1Pediatrics, 3Medicine, 4Anatomy, Cell Biology & Physiology, 5Biochemistry & Molecular Biology, the 6Center for Diabetes & Metabolic Diseases, and the 7Herman B. Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202; 2Department of BioHealth Informatics, Indiana University-Purdue University Indianapolis, Indianapolis, IN, 46202; 8Roudebush VA Medical Center, Indianapolis, IN 46202. *Corresponding Author(s): Carmella Evans-Molina, MD, PhD ([email protected]) Indiana University School of Medicine, 635 Barnhill Drive, MS 2031A, Indianapolis, IN 46202, Telephone: (317) 274-4145, Fax (317) 274-4107 Running Title: Golgi Stress Response in Diabetes Word Count: 4358 Number of Figures: 6 Keywords: Golgi apparatus stress, Islets, β cell, Type 1 diabetes, Type 2 diabetes 1 Diabetes Publish Ahead of Print, published online August 20, 2020 Diabetes Page 2 of 781 ABSTRACT The Golgi apparatus (GA) is an important site of insulin processing and granule maturation, but whether GA organelle dysfunction and GA stress are present in the diabetic β-cell has not been tested. We utilized an informatics-based approach to develop a transcriptional signature of β-cell GA stress using existing RNA sequencing and microarray datasets generated using human islets from donors with diabetes and islets where type 1(T1D) and type 2 diabetes (T2D) had been modeled ex vivo. To narrow our results to GA-specific genes, we applied a filter set of 1,030 genes accepted as GA associated.
    [Show full text]
  • Material Data Sheet
    MATERIAL DATA SHEET Recombinant Human His6 UBE2N/UBE2V2 Complex Cat. # E2­666 Ubiquitin conjugating Enzyme E2N (UBE2N), also known as Ubiquitin conjugating Enzyme 13 (Ubc13), forms a functional complex with the catalytically inactive UBE2V2 (human homologue of yeast MMS2) protein (1). Human UBE2N/Ubc13 shares 100% and 99% amino acid (aa) sequence identity with the mouse and rat orthologs, respectively, while human UBE2V2 shares 99% aa sequence identity with its mouse and rat orthologs. The UBE2N/UBE2V2 Complex functions with Ubiquitin ligases (E3s), including RNF111 and RNF8, to synthesize Lys63­linked Ubiquitin chains (2,3) that can either be unanchored or attached to target proteins (4, 5). The UBE2N/UBE2V2 complex has important roles in facilitating responses to various forms of DNA damage (2, 6). Product Information Quantity: 100 µg | 50 µg MW: 18 kDa (UBE2N), 17 kDa (UBE2V2) Source: E. coli­derived Contains an N­terminal 6­His tag Accession # P61088 (UBE2N)/Q15819 (UBE2V2) Stock: 0.88 mg/ml (25 μM) in 50 mM HEPES pH 7.5, 200 mM NaCl, 10% Glycerol (v/v), 2 mM TCEP Purity: >95%, by SDS­PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain. Rev. 5/22/2014 Page 1 of 2 www.bostonbiochem.com Boston Biochem products are available via the R&D Systems distributor network. USA & CANADA Tel: (800) 343-7475 EUROPE Tel: +44 (0)1235 529449 CHINA Tel: +86 (21) 52380373 Use & Storage Use: Recombinant Human His6­UBE2N/UBE2V2 Complex is a member of the Ubiquitin conjugating (E2) enzyme family that receives Ubiquitin from a Ubiquitin activating (E1) enzyme and subsequently interacts with a Ubiquitin ligase (E3) to conjugate Ubiquitin to substrate proteins.
    [Show full text]
  • Supplementary Tables
    Supplementary Tables Supplementary Table S1: Preselected miRNAs used in feature selection Univariate Cox proportional hazards regression analysis of the endpoint freedom from recurrence in the training set (DKTK-ROG sample) allowed the pre-selection of 524 miRNAs (P< 0.5), which were used in the feature selection. P-value was derived from log-rank test. miRNA p-value miRNA p-value miRNA p-value miRNA p-value hsa-let-7g-3p 0.0001520 hsa-miR-1304-3p 0.0490161 hsa-miR-7108-5p 0.1263245 hsa-miR-6865-5p 0.2073121 hsa-miR-6825-3p 0.0004257 hsa-miR-4298 0.0506194 hsa-miR-4453 0.1270967 hsa-miR-6893-5p 0.2120664 hsa-miR-668-3p 0.0005188 hsa-miR-484 0.0518625 hsa-miR-200a-5p 0.1276345 hsa-miR-25-3p 0.2123829 hsa-miR-3622b-3p 0.0005885 hsa-miR-6851-3p 0.0531446 hsa-miR-6090 0.1278692 hsa-miR-3189-5p 0.2136060 hsa-miR-6885-3p 0.0006452 hsa-miR-1276 0.0557418 hsa-miR-148b-3p 0.1279811 hsa-miR-6073 0.2139702 hsa-miR-6875-3p 0.0008188 hsa-miR-3173-3p 0.0559962 hsa-miR-4425 0.1288330 hsa-miR-765 0.2141536 hsa-miR-487b-5p 0.0011381 hsa-miR-650 0.0564616 hsa-miR-6798-3p 0.1293342 hsa-miR-338-5p 0.2153079 hsa-miR-210-5p 0.0012316 hsa-miR-6133 0.0571407 hsa-miR-4472 0.1300006 hsa-miR-6806-5p 0.2173515 hsa-miR-1470 0.0012822 hsa-miR-4701-5p 0.0571720 hsa-miR-4465 0.1304841 hsa-miR-98-5p 0.2184947 hsa-miR-6890-3p 0.0016539 hsa-miR-202-3p 0.0575741 hsa-miR-514b-5p 0.1308790 hsa-miR-500a-3p 0.2185577 hsa-miR-6511b-3p 0.0017165 hsa-miR-4733-5p 0.0616138 hsa-miR-378c 0.1317442 hsa-miR-4515 0.2187539 hsa-miR-7109-3p 0.0021381 hsa-miR-595 0.0629350 hsa-miR-3121-3p
    [Show full text]
  • Developmental Adaptations of Energy and Lipid Metabolism in Trypanosoma Brucei Insect Forms
    Department Biologie I Bereich Genetik Ludwig-Maximilians-Universität München Developmental adaptations of energy and lipid metabolism in Trypanosoma brucei insect forms. Stefan Allmann Dissertation der Fakultät für Biologie, Ludwig-Maximilians-Universität München Eingereicht am 18.09.2014 Erster Gutachter : Prof. Dr. Michael Boshart Biozentrum der Ludwig-Maximilians-Universität München Bereich Genetik Zweiter Gutachter: Prof. Dr. Peter Geigenberger Biozentrum der Ludwig-Maximilians-Universität München Bereich Pflanzenwissenschaften Datum der Abgabe: 18.09.2014 Datum der mündlichen Prüfung: 24.10.2014 Eidesstattliche Erklärung Ich versichere hiermit an Eides statt, dass die vorgelegte Dissertation von mir selbstständig und ohne unerlaubte Hilfe angefertigt wurde. Des Weiteren erkläre ich, dass ich nicht anderweitig ohne Erfolg versucht habe eine Dissertation einzureichen oder mich der Doktorprüfung zu unterziehen. Die folgende Dissertation liegt weder ganz, noch in wesentlichen Teilen einer anderen Prüfungskommission vor. München, 18.09.2014 Statutory declaration I declare that I have authored this thesis independently, that I have not used other than the declared sources/resources. As well I declare, that I have not submitted a dissertation without success and not passed the oral exam. The present dissertation (neither the entire dissertation nor parts) has not been presented to another examination board. Munich, 18.09.2014 III Content Eidesstattliche Erklärung III Statutory declaration III Abbreviations V Publications and Manuscripts Originating from this Thesis VII Contribution to Publications and Manuscripts Presented in this Thesis VIII Summary IX Zusammenfassung X 1. Introduction 11 1.1 Trypanosoma brucei in a nutshell 11 1.2 The parasite’s life cycle 12 1.3 Intermediate and energy metabolism 14 1.4 NADPH balance 17 1.5 Lipid droplets as energy storage 20 2.
    [Show full text]
  • WO 2019/079361 Al 25 April 2019 (25.04.2019) W 1P O PCT
    (12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property Organization I International Bureau (10) International Publication Number (43) International Publication Date WO 2019/079361 Al 25 April 2019 (25.04.2019) W 1P O PCT (51) International Patent Classification: CA, CH, CL, CN, CO, CR, CU, CZ, DE, DJ, DK, DM, DO, C12Q 1/68 (2018.01) A61P 31/18 (2006.01) DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, C12Q 1/70 (2006.01) HR, HU, ID, IL, IN, IR, IS, JO, JP, KE, KG, KH, KN, KP, KR, KW, KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, (21) International Application Number: MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, PCT/US2018/056167 OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, (22) International Filing Date: SC, SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, 16 October 2018 (16. 10.2018) TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW. (25) Filing Language: English (84) Designated States (unless otherwise indicated, for every kind of regional protection available): ARIPO (BW, GH, (26) Publication Language: English GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, SZ, TZ, (30) Priority Data: UG, ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, RU, TJ, 62/573,025 16 October 2017 (16. 10.2017) US TM), European (AL, AT, BE, BG, CH, CY, CZ, DE, DK, EE, ES, FI, FR, GB, GR, HR, HU, ΓΕ , IS, IT, LT, LU, LV, (71) Applicant: MASSACHUSETTS INSTITUTE OF MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, SM, TECHNOLOGY [US/US]; 77 Massachusetts Avenue, TR), OAPI (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, GW, Cambridge, Massachusetts 02139 (US).
    [Show full text]
  • The UBE2L3 Ubiquitin Conjugating Enzyme: Interplay with Inflammasome Signalling and Bacterial Ubiquitin Ligases
    The UBE2L3 ubiquitin conjugating enzyme: interplay with inflammasome signalling and bacterial ubiquitin ligases Matthew James George Eldridge 2018 Imperial College London Department of Medicine Submitted to Imperial College London for the degree of Doctor of Philosophy 1 Abstract Inflammasome-controlled immune responses such as IL-1β release and pyroptosis play key roles in antimicrobial immunity and are heavily implicated in multiple hereditary autoimmune diseases. Despite extensive knowledge of the mechanisms regulating inflammasome activation, many downstream responses remain poorly understood or uncharacterised. The cysteine protease caspase-1 is the executor of inflammasome responses, therefore identifying and characterising its substrates is vital for better understanding of inflammasome-mediated effector mechanisms. Using unbiased proteomics, the Shenoy grouped identified the ubiquitin conjugating enzyme UBE2L3 as a target of caspase-1. In this work, I have confirmed UBE2L3 as an indirect target of caspase-1 and characterised its role in inflammasomes-mediated immune responses. I show that UBE2L3 functions in the negative regulation of cellular pro-IL-1 via the ubiquitin- proteasome system. Following inflammatory stimuli, UBE2L3 assists in the ubiquitylation and degradation of newly produced pro-IL-1. However, in response to caspase-1 activation, UBE2L3 is itself targeted for degradation by the proteasome in a caspase-1-dependent manner, thereby liberating an additional pool of IL-1 which may be processed and released. UBE2L3 therefore acts a molecular rheostat, conferring caspase-1 an additional level of control over this potent cytokine, ensuring that it is efficiently secreted only in appropriate circumstances. These findings on UBE2L3 have implications for IL-1- driven pathology in hereditary fever syndromes, and autoinflammatory conditions associated with UBE2L3 polymorphisms.
    [Show full text]
  • Characterization of the Cellular Network of Ubiquitin Conjugating and Ligating Enzymes Ewa Katarzyna Blaszczak
    Characterization of the cellular network of ubiquitin conjugating and ligating enzymes Ewa Katarzyna Blaszczak To cite this version: Ewa Katarzyna Blaszczak. Characterization of the cellular network of ubiquitin conjugating and ligating enzymes. Cellular Biology. Université Rennes 1, 2015. English. NNT : 2015REN1S116. tel-01547616 HAL Id: tel-01547616 https://tel.archives-ouvertes.fr/tel-01547616 Submitted on 27 Jun 2017 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. ANNÉE 2015 THÈSE / UNIVERSITÉ DE RENNES 1 sous le sceau de l’Université Européenne de Bretagne pour le grade de DOCTEUR DE L’UNIVERSITÉ DE RENNES 1 Mention : BIOLOGIE École doctorale Vie-Agro-Santé présentée par Ewa Katarzyna Blaszczak Préparée à l’unité de recherche UMR 6290, IGDR Institut de Génétique et Développement de Rennes Université Rennes 1 Thèse soutenue à Rennes le 26.06.2015 Characterization of devant le jury composé de : Aude ECHALIER-GLAZER the cellular network Maître de conférence University of Leicester / rapporteur of ubiquitin Lionel PINTARD Directeur de recherche
    [Show full text]
  • ABSTRACT Title of Dissertation
    ABSTRACT Title of Dissertation: COMPARATIVE TRANSCRIPTOME PROFILING OF HUMAN FORESKIN FIBROBLASTS INFECTED WITH THE SYLVIO AND Y STRAINS OF TRYPANOSOMA CRUZI Genevieve Houston-Ludlam, Doctor of Philosophy, 2016 Dissertation Directed by: Professor Najib M. El-Sayed Department of Cell Biology and Molecular Genetics Trypanosoma cruzi, the causative agent of Chagas Disease, is phylogenetically distributed into nearly identical genetic strains which show divergent clinical presentations including differences in rates of cardiomyopathy in humans, different vector species and transmission cycles, and differential congenital transmission in a mouse model. The population structure of these strains divides into two groups, which are geographically and clinically distinct. The aim of this study was to compare the transcriptome of two strains of T. cruzi, Sylvio vs. Y to identify differences in expression that could account for clinical and biochemical differences. We collected and sequenced RNA from T. cruzi-infected and control Human Foreskin Fibroblasts at three timepoints. Differential expression analysis identified gene expression profiles at different timepoints in Sylvio infections, and between Sylvio and Y infections in both parasite and host. The Sylvio strain parasite and the host response to Sylvio infection largely mirrored the host- pathogen interaction seen in our previous Y strain work. IL-8 was more highly expressed in Sylvio-infected HFFs than in Y-infected HFFs. Comparative transcriptome profiling of human foreskin fibroblasts infected with the Sylvio and Y strains of Trypanosoma cruzi By Genevieve Houston-Ludlam Dissertation submitted to the Faculty of the Graduate School of the University of Maryland, College Park, in partial fulfillment of the requirements for the degree of Doctor of Philosophy 2016 Advisory Committee: Professor Najib M.
    [Show full text]
  • Dissertation.Pdf
    A Cluster of Antimony Resistance Genes on Chromosome 34 of Leishmania infantum and Their Properties Dissertation with the aim of achieving a doctoral degree at the Faculty of Mathematics, Informatics and Natural Sciences Department of Biology of Universität Hamburg Submitted by Paloma Tejera Nevado 2016 Hamburg This work has been performed from May 2013 to April 2016 in the research group of PD Dr. Joachim Clos at the Bernhard-Nocht-Institute for Tropical Medicine in Hamburg. 1. Evaluator: Prof. Dr. Wilhelm Schäfer Biozentrum Klein Flottbek Abteilung für Molekulare Phytopathologie und Genetik Ohnhorstst. 18, 22609 Hamburg 2. Evaluator: PD Dr. Joachim Clos Bernhard-Nocht-Institut für Tropenmedizin Abteilung für Molekulare Parasitologie Bernhard-Nocht-Straße 74, 20359 Hamburg Day of oral defense: 15th July 2016 Hiermit erkläre ich an Eides statt, dass ich die vorliegende Dissertationsschrift selbst verfasst und keine anderen als die angegebenen Quellen und Hilfsmittel benutzt habe. I hereby declare, on oath, that I have written the present dissertation by my own and have not used other than the acknowledged resources and aids. Hamburg, 2016 Signature Paloma Tejera Nevado Acknowledgements This thesis reflects part of the intensive work done during three years. During this time I have learnt a lot of things at the BNI. I would like to express my sincere gratitude to PD Dr Joachim Clos, who gave me the opportunity to do my doctoral studies in his lab. I would also like to thank my co supervisors at the institute PD Dr Thomas Jacobs and Dr Michael Schreiber and Prof. Dr Wihelm Schäfer at the UHH. I am especially grateful to my family, especially my parents and my sister.
    [Show full text]