DOCSLIB.ORG

Characterization of the Regulation of the Er Stress Response by the Dna Repair Enzyme Aag

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Characterization of the Regulation of the Er Stress Response by the Dna Repair Enzyme Aag DECIPHERING THE CROSSTALK: CHARACTERIZATION OF THE REGULATION OF THE ER STRESS RESPONSE BY THE DNA REPAIR ENZYME AAG Clara Forrer Charlier Faculty of Health and Medical Sciences Department of Biochemistry and Physiology This thesis is submitted for the degree of Doctor of Philosophy June 2018 DECIPHERING THE CROSSTALK: CHARACTERIZATION OF THE REGULATION OF THE ER STRESS RESPONSE BY THE DNA REPAIR ENZYME AAG- Clara Forrer Charlier – June 2018 SUMMARY The genome is a very dynamic store of genetic information and constantly threatened by endogenous and exogenous damaging agents. To maintain fidelity of the information stored, several robust and overlapping repair pathways, such as the Base Excision Repair (BER) pathway, have evolved. The main BER glycosylase responsible for repairing alkylation DNA damage is the alkyladenine DNA glycosylase (AAG). Repair initiated by AAG can lead to accumulation of cytotoxic intermediates. Here, we report the involvement of AAG in the elicitation of the unfolded protein response (UPR), a mechanism triggered to restore proteostasis in the cell whose dysfunction is implicated in diseases like diabetes, Alzheimer’s disease and cancer. Firstly, we determined that not only human ARPE-19 cells were capable of eliciting the UPR, but that an alkylating agent, methyl methanesulfonate (MMS), also triggers the response, and that in the absence of AAG the response is greatly diminished. Our luciferase reporter assay indicates that the response is activated on multiple branches (IRE1 and ATF6) on both AAG-proficient and deficient cells. Although no transcriptional induction of UPR markers was detected by RT-qPCR at 6 hours post MMS treatment, preliminary western-blot data at 6 and 24h, show activation of key UPR markers (p-eIF2α, BiP and XBP-1) upon MMS treatment in wild-type cells and little or no activation on AAG -/-. To investigate the impact of AAG modulation on the cellular proteome we also conducted a proteomic analysis, identifying 5480 protein groups in wild-type ARPE-19, 5377 in the AAG -/- A2C2 and 5264 in AAG -/- B6C3. After a high-stringency analysis, we identified 13 proteins present only in wild-type cells, indicating promising targets for further investigation into the role of AAG in the UPR. We also identified 44 overrepresented GO-slim terms across all cell lines and 23 overrepresented pathways, mostly related to cellular metabolism processes. Whereas more experiments are required to characterize the nature of AAG’s contribution to the UPR, we demonstrate the existence of crosstalk between the DNA repair response and the ER stress response pathways, that is potentially relevant in a clinical setting. ii ACKNOWLEDGEMENTS There is always a little trepidation whenever one needs to write acknowledgements. It was such a long and intense journey, meeting so many wonderful people that is hard not to feel that you are missing someone important in this section. Firstly, I would like to thank my mother, without whom I wouldn’t be here, both literally and figuratively. Thanks for being such an incredibly wise person, thanks for your support, patience and love. Thanks for stimulating my scientific curiosity and for believing in me. You are my first and best friend, always. And thanks for the mitochondria! I like to thank Lisi, for also believing in me and letting me be a part of her fantastic group. You are not only a fantastic supervisor, but a fantastic human being as well. Every time I replied to somebody saying you were my supervisor I invariably heard back “she is lovely!” or “you are very lucky!”. I know I am. You rock! You are funny, supportive and the owner of a very sharp mind. I have grown as much as a person as a scientist after I met you. Don’t ever change! Have I mentioned you are also a great cook and have the most contagious laughter I ever heard? Thanks for having me! Thanks to Bernadette Moore for all the pep talk, writing tips and for introducing me to the scary world of proteomics. Although my first brush with it was a bit rocky, I am always keen to learn something completely new. Thanks for constantly worrying about my sanity as well! Thanks to Axel Nohturfft for helping (a lot!) with the proteomic data analysis, the plasmids, the cells and for being the husband of the best supervisor ever. iii Thanks to Dr Ruan Elliott and Eleanor Healing for developing the AAG activity assay and applying the AAG activity assay on the bazillions of overexpressors, wild-types and knockout clones I had. Thanks to Dr Paul Skipp and Dr Erika Parkinson for having me on the Proteomics Lab at the University of Southampton. Thanks for enduring all my silly questions with patience and grace (i.e. not laughing at them) and for letting me use the sexiest and most expensive piece of equipment I ever touched. Thanks to Dr J. A. Pêgas Henriques, for the all the support back in Brazil (bureaucratic and otherwise) and for putting me in touch with Lisi. Thanks to Dr Izabel Vilella for encouraging me to embark in the adventure that was my PhD here in England. It wouldn’t have happened if not for you. Thanks for all the fantastic people I met on this journey, specially my colleagues from 04AY04: Larissa Milano, for the help on the lab, for listening to me moan since my Masters and for being the only other fellow coffee drinker in the office. And for helping me survive a whole week eating only salmon on all meals at Egmond aan Zee. Thanks Natalie Mayhead for all the fun, signing, booze, science, Christmases and nerdy stuff and for putting up with my weirdness and grumpiness. Thanks even for the occasional hug (ewww!). Thanks Abdullah Aljohani for being such a great colleague and for sharing my love for good food. Thanks for all the recipes and all the laughter we had together. Thank you and Fahad Alhumaydhi for the warm reception I got when I first arrived, wide-eyed and really scared. Go Team AAG! Thanks to our extended lab family: the lovely and smart Amy Barber (although she never gave me the pictures of our last lab Christmas party), the stylish Matshediso Zachariah (one day I’ll drop by to camp with you and see the lions you promised), the graceful Diana Bordin (now gazing at the northern lights) and grumpy Leoncikas Vytautas (former colleague and Singstar Addict). I am eternally iv happy for being part of your lives and I am going to miss you a lot. Maybe I’ll even cry a little (but just a little). Hidden somewhere, probably, so I don’t ruin my reputation. Love you all! Thanks for the CNPq/Science Without Borders for giving this priceless opportunity (and paying me on time). It was a childhood dream come true. Finally, thanks for all the people that helped make this happen in some way, being with technical advice, food, alcohol, laughter or for lending me an ear when I needed. Sorry if I forgot to mention you here! v CONTENTS 1 INTRODUCTION .................................................................................... 25 DNA DAMAGE AND REPAIR ....................................................................... 25 1.1.1 Lesions repaired by BER ............................................................ 27 1.1.2 Alkyladenine DNA Glycosylase/Methylpurine DNA Glycosylase (AAG/MPG) ............................................................ 41 ER STRESS AND UNFOLDED PROTEIN RESPONSE ............................................. 52 1.2.1 IRE1 ........................................................................................... 54 1.2.2 PRKR-Like Endoplasmic Reticulum Kinase - PERK ..................... 58 1.2.3 Activating Transcription Factor 6 .............................................. 63 1.2.4 Endoplasmic Reticulum-Associated Degradation - ERAD ......... 65 PROBLEM STATEMENT AND OBJECTIVES ........................................................ 69 2 EVALUATION OF THE PHENOTYPES RESULTING FROM AAG EXPRESSION IN DIFFERENT SUBCELLULAR COMPARTMENTS ................. 71 INTRODUCTION ........................................................................................ 71 AIM ....................................................................................................... 74 OBJECTIVES ............................................................................................. 74 MATERIALS AND METHODS ........................................................................ 74 2.4.1 Cloning strategies ...................................................................... 74 2.4.2 Cell culture ................................................................................. 78 2.4.3 Isogenic cell line generation ...................................................... 79 2.4.4 Cell line characterization ........................................................... 79 RESULTS ................................................................................................. 84 2.5.1 Generation and characterization of the AAG overexpressing isogenic cell lines ....................................................................... 84 2.5.2 AAG overexpression sensitizes cells to alkylating damage ....... 86 2.5.3 AAG knockdown seems to decrease cell fitness ........................ 87 DISCUSSION ............................................................................................ 89 3 AAG AND THE UPR: CELL LINE DEVELOPMENT AND CHARACTERIZATION ............................................................................. 92 vi INTRODUCTION ........................................................................................ 92 AIMS ....................................................................................................
Load more

Recommended publications
  • BER and Folate Deficiency: a Comprehensive Overview of DNA Base Excision Repair and the Effect of Dietary Folate Lydia Lanni [email protected]
    Wayne State University Honors College Theses Irvin D. Reid Honors College Winter 5-7-2012 BER and Folate Deficiency: A comprehensive overview of DNA base excision repair and the effect of dietary folate Lydia Lanni [email protected] Follow this and additional works at: https://digitalcommons.wayne.edu/honorstheses Part of the Human and Clinical Nutrition Commons Recommended Citation Lanni, Lydia, "BER and Folate Deficiency: A comprehensive overview of DNA base excision repair and the effect of dietary folate" (2012). Honors College Theses. 2. https://digitalcommons.wayne.edu/honorstheses/2 This Honors Thesis is brought to you for free and open access by the Irvin D. Reid Honors College at DigitalCommons@WayneState. It has been accepted for inclusion in Honors College Theses by an authorized administrator of DigitalCommons@WayneState. Running head: BER AND FOLATE DEFICIENCY 1 BER and Folate Deficiency: A comprehensive overview of DNA base excision repair and the effect of dietary folate Lydia Lanni Wayne State University Honors Thesis Winter 2012 Author Note A special thank you to Dr. Diane Cabelof and all those in her lab for their instruction and support. BER AND FOLATE DEFICIENCY 2 Abstract Folate is the naturally occurring form of water-soluble B vitamin that is found in foods such as leafy vegetables, fruits, legumes, etc. Dietary supplementation of folate has shown to be protective against neural tube defects and other congenital disorders, and of recent, its role in carcinogenesis has been of special interest. Though mechanistically unclear, a positive correlation has been observed between folate deficiency in the diet and decrease function of DNA base excision repair pathways.
    [Show full text]
  • Computational Genome-Wide Identification of Heat Shock Protein Genes in the Bovine Genome [Version 1; Peer Review: 2 Approved, 1 Approved with Reservations]
    F1000Research 2018, 7:1504 Last updated: 08 AUG 2021 RESEARCH ARTICLE Computational genome-wide identification of heat shock protein genes in the bovine genome [version 1; peer review: 2 approved, 1 approved with reservations] Oyeyemi O. Ajayi1,2, Sunday O. Peters3, Marcos De Donato2,4, Sunday O. Sowande5, Fidalis D.N. Mujibi6, Olanrewaju B. Morenikeji2,7, Bolaji N. Thomas 8, Matthew A. Adeleke 9, Ikhide G. Imumorin2,10,11 1Department of Animal Breeding and Genetics, Federal University of Agriculture, Abeokuta, Nigeria 2International Programs, College of Agriculture and Life Sciences, Cornell University, Ithaca, NY, 14853, USA 3Department of Animal Science, Berry College, Mount Berry, GA, 30149, USA 4Departamento Regional de Bioingenierias, Tecnologico de Monterrey, Escuela de Ingenieria y Ciencias, Queretaro, Mexico 5Department of Animal Production and Health, Federal University of Agriculture, Abeokuta, Nigeria 6Usomi Limited, Nairobi, Kenya 7Department of Animal Production and Health, Federal University of Technology, Akure, Nigeria 8Department of Biomedical Sciences, Rochester Institute of Technology, Rochester, NY, 14623, USA 9School of Life Sciences, University of KwaZulu-Natal, Durban, 4000, South Africa 10School of Biological Sciences, Georgia Institute of Technology, Atlanta, GA, 30032, USA 11African Institute of Bioscience Research and Training, Ibadan, Nigeria v1 First published: 20 Sep 2018, 7:1504 Open Peer Review https://doi.org/10.12688/f1000research.16058.1 Latest published: 20 Sep 2018, 7:1504 https://doi.org/10.12688/f1000research.16058.1 Reviewer Status Invited Reviewers Abstract Background: Heat shock proteins (HSPs) are molecular chaperones 1 2 3 known to bind and sequester client proteins under stress. Methods: To identify and better understand some of these proteins, version 1 we carried out a computational genome-wide survey of the bovine 20 Sep 2018 report report report genome.
    [Show full text]
  • Genomic Correlates of Relationship QTL Involved in Fore- Versus Hind Limb Divergence in Mice
    Loyola University Chicago Loyola eCommons Biology: Faculty Publications and Other Works Faculty Publications 2013 Genomic Correlates of Relationship QTL Involved in Fore- Versus Hind Limb Divergence in Mice Mihaela Palicev Gunter P. Wagner James P. Noonan Benedikt Hallgrimsson James M. Cheverud Loyola University Chicago, [email protected] Follow this and additional works at: https://ecommons.luc.edu/biology_facpubs Part of the Biology Commons Recommended Citation Palicev, M, GP Wagner, JP Noonan, B Hallgrimsson, and JM Cheverud. "Genomic Correlates of Relationship QTL Involved in Fore- Versus Hind Limb Divergence in Mice." Genome Biology and Evolution 5(10), 2013. This Article is brought to you for free and open access by the Faculty Publications at Loyola eCommons. It has been accepted for inclusion in Biology: Faculty Publications and Other Works by an authorized administrator of Loyola eCommons. For more information, please contact [email protected]. This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 License. © Palicev et al., 2013. GBE Genomic Correlates of Relationship QTL Involved in Fore- versus Hind Limb Divergence in Mice Mihaela Pavlicev1,2,*, Gu¨ nter P. Wagner3, James P. Noonan4, Benedikt Hallgrı´msson5,and James M. Cheverud6 1Konrad Lorenz Institute for Evolution and Cognition Research, Altenberg, Austria 2Department of Pediatrics, Cincinnati Children‘s Hospital Medical Center, Cincinnati, Ohio 3Yale Systems Biology Institute and Department of Ecology and Evolutionary Biology, Yale University 4Department of Genetics, Yale University School of Medicine 5Department of Cell Biology and Anatomy, The McCaig Institute for Bone and Joint Health and the Alberta Children’s Hospital Research Institute for Child and Maternal Health, University of Calgary, Calgary, Canada 6Department of Anatomy and Neurobiology, Washington University *Corresponding author: E-mail: [email protected].
    [Show full text]
  • Oxidative Stress in Sperm Affects the Epigenetic Reprogramming in Early
    Wyck et al. Epigenetics & Chromatin (2018) 11:60 https://doi.org/10.1186/s13072-018-0224-y Epigenetics & Chromatin RESEARCH Open Access Oxidative stress in sperm afects the epigenetic reprogramming in early embryonic development Sarah Wyck1,2,3, Carolina Herrera1, Cristina E. Requena4,5, Lilli Bittner1, Petra Hajkova4,5, Heinrich Bollwein1* and Rafaella Santoro2* Abstract Background: Reactive oxygen species (ROS)-induced oxidative stress is well known to play a major role in male infer- tility. Sperm are sensitive to ROS damaging efects because as male germ cells form mature sperm they progressively lose the ability to repair DNA damage. However, how oxidative DNA lesions in sperm afect early embryonic develop- ment remains elusive. Results: Using cattle as model, we show that fertilization using sperm exposed to oxidative stress caused a major developmental arrest at the time of embryonic genome activation. The levels of DNA damage response did not directly correlate with the degree of developmental defects. The early cellular response for DNA damage, γH2AX, is already present at high levels in zygotes that progress normally in development and did not signifcantly increase at the paternal genome containing oxidative DNA lesions. Moreover, XRCC1, a factor implicated in the last step of base excision repair (BER) pathway, was recruited to the damaged paternal genome, indicating that the maternal BER machinery can repair these DNA lesions induced in sperm. Remarkably, the paternal genome with oxidative DNA lesions showed an impairment of zygotic active DNA demethylation, a process that previous studies linked to BER. Quantitative immunofuorescence analysis and ultrasensitive LC–MS-based measurements revealed that oxidative DNA lesions in sperm impair active DNA demethylation at paternal pronuclei, without afecting 5-hydroxymethyl- cytosine (5hmC), a 5-methylcytosine modifcation that has been implicated in paternal active DNA demethylation in mouse zygotes.
    [Show full text]
  • Gene Targeting Therapies (Roy Alcalay)
    Recent Developments in Gene - Targeted Therapies for Parkinson’s Disease Roy Alcalay, MD, MS Alfred and Minnie Bressler Associate Professor of Neurology Division of Movement Disorders Columbia University Medical Center Disclosures Funding: Dr. Alcalay is funded by the National Institutes of Health, the DOD, the Michael J. Fox Foundation and the Parkinson’s Foundation. Dr. Alcalay receives consultation fees from Genzyme/Sanofi, Restorbio, Janssen, and Roche. Gene Localizations Identified in PD Gene Symbol Protein Transmission Chromosome PARK1 SNCA α-synuclein AD 4q22.1 PARK2 PRKN parkin (ubiquitin ligase) AR 6q26 PARK3 ? ? AD 2p13 PARK4 SNCA triplication α-synuclein AD 4q22.1 PARK5 UCH-L1 ubiquitin C-terminal AD 4p13 hydrolase-L1 PARK6 PINK1 PTEN-induced kinase 1 AR 1p36.12 PARK7 DJ-1 DJ-1 AR 1p36.23 PARK8 LRRK2 leucine rich repeat kinase 2 AD 12q12 PARK9 ATP13A2 lysosomal ATPase AR 1p36.13 PARK10 ? ? (Iceland) AR 1p32 PARK11 GIGYF2 GRB10-interacting GYF protein 2 AD 2q37.1 PARK12 ? ? X-R Xq21-q25 PARK13 HTRA2 serine protease AD 2p13.1 PARK14 PLA2G6 phospholipase A2 (INAD) AR 22q13.1 PARK15 FBXO7 F-box only protein 7 AR 22q12.3 PARK16 ? Discovered by GWAS ? 1q32 PARK17 VPS35 vacuolar protein sorting 35 AD 16q11.2 PARK18 EIF4G1 initiation of protein synth AD 3q27.1 PARK19 DNAJC6 auxilin AR 1p31.3 PARK20 SYNJ1 synaptojanin 1 AR 21q22.11 PARK21 DNAJC13 8/RME-8 AD 3q22.1 PARK22 CHCHD2 AD 7p11.2 PARK23 VPS13C AR 15q22 Gene Localizations Identified in PD Disorder Symbol Protein Transmission Chromosome PD GBA β-glucocerebrosidase AD 1q21 SCA2
    [Show full text]
  • Table 2. Significant
    Table 2. Significant (Q < 0.05 and |d | > 0.5) transcripts from the meta-analysis Gene Chr Mb Gene Name Affy ProbeSet cDNA_IDs d HAP/LAP d HAP/LAP d d IS Average d Ztest P values Q-value Symbol ID (study #5) 1 2 STS B2m 2 122 beta-2 microglobulin 1452428_a_at AI848245 1.75334941 4 3.2 4 3.2316485 1.07398E-09 5.69E-08 Man2b1 8 84.4 mannosidase 2, alpha B1 1416340_a_at H4049B01 3.75722111 3.87309653 2.1 1.6 2.84852656 5.32443E-07 1.58E-05 1110032A03Rik 9 50.9 RIKEN cDNA 1110032A03 gene 1417211_a_at H4035E05 4 1.66015788 4 1.7 2.82772795 2.94266E-05 0.000527 NA 9 48.5 --- 1456111_at 3.43701477 1.85785922 4 2 2.8237185 9.97969E-08 3.48E-06 Scn4b 9 45.3 Sodium channel, type IV, beta 1434008_at AI844796 3.79536664 1.63774235 3.3 2.3 2.75319499 1.48057E-08 6.21E-07 polypeptide Gadd45gip1 8 84.1 RIKEN cDNA 2310040G17 gene 1417619_at 4 3.38875643 1.4 2 2.69163229 8.84279E-06 0.0001904 BC056474 15 12.1 Mus musculus cDNA clone 1424117_at H3030A06 3.95752801 2.42838452 1.9 2.2 2.62132809 1.3344E-08 5.66E-07 MGC:67360 IMAGE:6823629, complete cds NA 4 153 guanine nucleotide binding protein, 1454696_at -3.46081884 -4 -1.3 -1.6 -2.6026947 8.58458E-05 0.0012617 beta 1 Gnb1 4 153 guanine nucleotide binding protein, 1417432_a_at H3094D02 -3.13334396 -4 -1.6 -1.7 -2.5946297 1.04542E-05 0.0002202 beta 1 Gadd45gip1 8 84.1 RAD23a homolog (S.
    [Show full text]
  • Characterization of Α-L-Fucosidase and Other Digestive Hydrolases From
    Acta Tropica 141 (2015) 118–127 Contents lists available at ScienceDirect Acta Tropica journal homepage: www.elsevier.com/locate/actatropica Characterization of ␣-L-fucosidase and other digestive hydrolases from Biomphalaria glabrata Natalia N. Perrella a,b, Rebeca S. Cantinha c,d, Eliana Nakano c, Adriana R. Lopes a,∗ a Laboratory of Biochemistry and Biophysics—Instituto Butantan, São Paulo, Brazil b Programa de Pós Graduac¸ ão Interunidades em Biotecnologia PPIB, Universidade de São Paulo, São Paulo, SP, Brazil c Laboratory of Parasitology—Instituto Butantan, São Paulo, Brazil d Instituto de Pesquisas Energéticas e Nucleares, Universidade de São Paulo, São Paulo, SP, Brazil article info abstract Article history: Schistosoma mansoni is one of the major agents of the disease Schistosomiasis, which is one of the Received 10 February 2014 major global public health concerns. Biomphalaria glabrata is an obligate intermediate mollusc host of Received in revised form 3 July 2014 S. mansoni. Although the development of S. mansoni occurs in the snail hepatopancreas, studies that Accepted 12 August 2014 focus on this organ remain limited. In this study, we biochemically identified five distinct carbohy- Available online 16 September 2014 drases (amylase, maltase, ␣-glucosidase, trehalase, and ␣-L-fucosidase), lipases, and peptidases in the B. glabrata hepatopancreas and focused on the isolation and characterization of the activity of ␣-L- Keywords: fucosidase. The isolated ␣-L-fucosidase has a molecular mass of 141 kDa, an optimum pH of 5.8, and Hepatopancreas ␣ Enzymes is inhibited by Tris, fucose, and 1-deoxyfuconojirimycin. B. glabrata -L-fucosidase is an exoglycosidase ␮ ␣-L-Fucosidase that can hydrolyze the natural substrate fucoidan to fucose residues.
    [Show full text]
  • Characterisation of Streptococcus Pneumoniae Opacity Phase Variation
    Characterisation of Streptococcus pneumoniae Opacity Phase Variation Melissa Hui Chieh Chai, BBiomedSc (Hon), MSc A thesis submitted in fulfilment of the requirements for the degree of Doctor of Philosophy from the University of Adelaide February 2016 Research Centre for Infectious Diseases Department of Molecular and Cellular Biology The University of Adelaide Adelaide, S.A., Australia Table of Contents Chapter 1: INTRODUCTION ........................................................................... 1 1.1 The Pneumococcus ....................................................................................................... 1 1.1.1 Burden of Disease .................................................................................................. 1 1.1.2 Antimicrobial resistance ........................................................................................ 3 1.2 Pneumococcal Vaccines ............................................................................................... 4 1.3 Pathogenesis of Pneumococcal Disease ....................................................................... 7 1.3.1 Pneumococcal colonisation.................................................................................... 7 1.3.2 Progression to disease .......................................................................................... 10 1.4 S. pneumoniae Virulence Factors ............................................................................... 11 1.4.1 Pneumococcal capsule ........................................................................................
    [Show full text]
  • Glycoproteomics-Based Signatures for Tumor Subtyping and Clinical Outcome Prediction of High-Grade Serous Ovarian Cancer
    ARTICLE https://doi.org/10.1038/s41467-020-19976-3 OPEN Glycoproteomics-based signatures for tumor subtyping and clinical outcome prediction of high-grade serous ovarian cancer Jianbo Pan 1,2,3, Yingwei Hu1,3, Shisheng Sun 1,3, Lijun Chen1, Michael Schnaubelt1, David Clark1, ✉ Minghui Ao1, Zhen Zhang1, Daniel Chan1, Jiang Qian2 & Hui Zhang 1 1234567890():,; Inter-tumor heterogeneity is a result of genomic, transcriptional, translational, and post- translational molecular features. To investigate the roles of protein glycosylation in the heterogeneity of high-grade serous ovarian carcinoma (HGSC), we perform mass spectrometry-based glycoproteomic characterization of 119 TCGA HGSC tissues. Cluster analysis of intact glycoproteomic profiles delineates 3 major tumor clusters and 5 groups of intact glycopeptides. It also shows a strong relationship between N-glycan structures and tumor molecular subtypes, one example of which being the association of fucosylation with mesenchymal subtype. Further survival analysis reveals that intact glycopeptide signatures of mesenchymal subtype are associated with a poor clinical outcome of HGSC. In addition, we study the expression of mRNAs, proteins, glycosites, and intact glycopeptides, as well as the expression levels of glycosylation enzymes involved in glycoprotein biosynthesis pathways in each tumor. The results show that glycoprotein levels are mainly controlled by the expression of their individual proteins, and, furthermore, that the glycoprotein-modifying glycans cor- respond to the protein levels of glycosylation enzymes. The variation in glycan types further shows coordination to the tumor heterogeneity. Deeper understanding of the glycosylation process and glycosylation production in different subtypes of HGSC may provide important clues for precision medicine and tumor-targeted therapy.
    [Show full text]
  • Dual Proteome-Scale Networks Reveal Cell-Specific Remodeling of the Human Interactome
    bioRxiv preprint doi: https://doi.org/10.1101/2020.01.19.905109; this version posted January 19, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Dual Proteome-scale Networks Reveal Cell-specific Remodeling of the Human Interactome Edward L. Huttlin1*, Raphael J. Bruckner1,3, Jose Navarrete-Perea1, Joe R. Cannon1,4, Kurt Baltier1,5, Fana Gebreab1, Melanie P. Gygi1, Alexandra Thornock1, Gabriela Zarraga1,6, Stanley Tam1,7, John Szpyt1, Alexandra Panov1, Hannah Parzen1,8, Sipei Fu1, Arvene Golbazi1, Eila Maenpaa1, Keegan Stricker1, Sanjukta Guha Thakurta1, Ramin Rad1, Joshua Pan2, David P. Nusinow1, Joao A. Paulo1, Devin K. Schweppe1, Laura Pontano Vaites1, J. Wade Harper1*, Steven P. Gygi1*# 1Department of Cell Biology, Harvard Medical School, Boston, MA, 02115, USA. 2Broad Institute, Cambridge, MA, 02142, USA. 3Present address: ICCB-Longwood Screening Facility, Harvard Medical School, Boston, MA, 02115, USA. 4Present address: Merck, West Point, PA, 19486, USA. 5Present address: IQ Proteomics, Cambridge, MA, 02139, USA. 6Present address: Vor Biopharma, Cambridge, MA, 02142, USA. 7Present address: Rubius Therapeutics, Cambridge, MA, 02139, USA. 8Present address: RPS North America, South Kingstown, RI, 02879, USA. *Correspondence: [email protected] (E.L.H.), [email protected] (J.W.H.), [email protected] (S.P.G.) #Lead Contact: [email protected] bioRxiv preprint doi: https://doi.org/10.1101/2020.01.19.905109; this version posted January 19, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder.
    [Show full text]
  • Glucose-Induced Changes in Gene Expression in Human Pancreatic Islets: Causes Or Consequences of Chronic Hyperglycemia
    Diabetes Volume 66, December 2017 3013 Glucose-Induced Changes in Gene Expression in Human Pancreatic Islets: Causes or Consequences of Chronic Hyperglycemia Emilia Ottosson-Laakso,1 Ulrika Krus,1 Petter Storm,1 Rashmi B. Prasad,1 Nikolay Oskolkov,1 Emma Ahlqvist,1 João Fadista,2 Ola Hansson,1 Leif Groop,1,3 and Petter Vikman1 Diabetes 2017;66:3013–3028 | https://doi.org/10.2337/db17-0311 Dysregulation of gene expression in islets from patients In patients with type 2 diabetes (T2D), islet function de- with type 2 diabetes (T2D) might be causally involved clines progressively. Although the initial pathogenic trigger in the development of hyperglycemia, or it could develop of impaired b-cell function is still unknown, elevated glu- as a consequence of hyperglycemia (i.e., glucotoxicity). cose levels are known to further aggravate b-cell function, a To separate the genes that could be causally involved condition referred to as glucotoxicity, which can stimulate in pathogenesis from those likely to be secondary to hy- apoptosis and lead to reduced b-cell mass (1–5). Prolonged perglycemia, we exposed islets from human donors to exposure to hyperglycemia also can induce endoplasmic re- ISLET STUDIES normal or high glucose concentrations for 24 h and ana- ticulum (ER) stress and production of reactive oxygen spe- fi lyzed gene expression. We compared these ndings with cies (6), which can further impair islet function and thereby gene expression in islets from donors with normal glucose the ability of islets to secrete the insulin needed to meet the tolerance and hyperglycemia (including T2D). The genes increased demands imposed by insulin resistance and obe- whose expression changed in the same direction after sity (7).
    [Show full text]
  • Fine Mapping Studies of Quantitative Trait Loci for Baseline Platelet Count in Mice and Humans
    Fine mapping studies of quantitative trait loci for baseline platelet count in mice and humans A thesis submitted in fulfillment of the requirements for the degree of Doctor of Philosophy Melody C Caramins December 2010 University of New South Wales ORIGINALITY STATEMENT ‘I hereby declare that this submission is my own work and to the best of my knowledge it contains no materials previously published or written by another person, or substantial proportions of material which have been accepted for the award of any other degree or diploma at UNSW or any other educational institution, except where due acknowledgement is made in the thesis. Any contribution made to the research by others, with whom I have worked at UNSW or elsewhere, is explicitly acknowledged in the thesis. I also declare that the intellectual content of this thesis is the product of my own work, except to the extent that assistance from others in the project's design and conception or in style, presentation and linguistic expression is acknowledged.’ Signed …………………………………………….............. Date …………………………………………….............. This thesis is dedicated to my father. Dad, thanks for the genes – and the environment! ACKNOWLEDGEMENTS “Nothing can come out of nothing, any more than a thing can go back to nothing.” - Marcus Aurelius Antoninus A PhD thesis is never the work of one person in isolation from the world at large. I would like to thank the following people, without whom this work would not have existed. Thank you firstly, to all my teachers, of which there have been many. Undoubtedly, the greatest debt is owed to my supervisor, Dr Michael Buckley.
    [Show full text]