DOCSLIB.ORG

Complete Dissertation

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Complete Dissertation VU Research Portal Molecular insight into the pathogenetic synergy between E. coli and B. fragilis in secondary peritonitis Sijbrandi, R. 2006 document version Publisher's PDF, also known as Version of record Link to publication in VU Research Portal citation for published version (APA) Sijbrandi, R. (2006). Molecular insight into the pathogenetic synergy between E. coli and B. fragilis in secondary peritonitis. General rights Copyright and moral rights for the publications made accessible in the public portal are retained by the authors and/or other copyright owners and it is a condition of accessing publications that users recognise and abide by the legal requirements associated with these rights. • Users may download and print one copy of any publication from the public portal for the purpose of private study or research. • You may not further distribute the material or use it for any profit-making activity or commercial gain • You may freely distribute the URL identifying the publication in the public portal ? Take down policy If you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediately and investigate your claim. E-mail address: [email protected] Download date: 09. Oct. 2021 MOLECULAR INSIGHT INTO THE PATHOGENIC SYNERGY BETWEEN E. COLI AND B. FRAGILIS IN SECONDARY PERITONITIS This thesis has been reviewed by: dr. W. Bitter, VU medisch centrum, Amsterdam dr. K. Bosscha, Jeroen Bosch Ziekenhuis, 's Hertogenbosch dr. E.N.G. Houben, University of Basel, Basel, Switzerland dr. L. Rutten, Universiteit Utrecht, Utrecht prof.dr. J.R.H. Tame, Yokohama City University, Yokohama, Japan dr. J.P. van Ulsen, Universiteit Utrecht, Utrecht Printed by Wöhrmann Print Service, Zutphen The studies presented in this thesis were carried out at the Department of Molecular Microbiology at the Vrije Universiteit Amsterdam in cooperation with the Genetics and Biochemistry Branch at the National Institutes of Health (Bethesda, MD, USA), the Protein Design Laboratory at the Yokohama City University (Yokohama, Japan), the Department of Crystal and Structural Chemistry and the Department of Molecular Microbiology at the Universiteit Utrecht, and the Department of Molecular Cytology at the Universiteit van Amsterdam. VRIJE UNIVERSITEIT Molecular insight into the pathogenic synergy between E. coli and B. fragilis in secondary peritonitis ACADEMISCH PROEFSCHRIFT ter verkrijging van de graad Doctor aan de Vrije Universiteit Amsterdam, op gezag van de rector magnificus prof.dr. T. Sminia, in het openbaar te verdedigen ten overstaan van de promotiecommissie van de faculteit der Aard- en Levenswetenschappen op maandag 27 maart 2006 om 13.45 uur in de aula van de universiteit, De Boelelaan 1105 door Robert Sijbrandi geboren te Ermelo promotor: prof.dr. B. Oudega copromotoren: dr. B.R. Otto dr. J. Luirink Contents Chapter 1 General introduction 7 Chapter 2 Signal Recognition Particle (SRP)-mediated targeting and Sec- 37 dependent translocation of an extracellular Escherichia coli protein Chapter 3 Crystal structure of Hemoglobin protease, a heme binding 55 autotransporter protein from pathogenic Escherichia coli Chapter 4 Purification, folding and preliminary structure of the translocator 71 domain of the Escherichia coli autotransporter Hbp Chapter 5 Characterization of an iron-regulated α-enolase of 89 Bacteroides fragilis Chapter 6 Cloning and preliminary characterization of Pbp, a putative 109 plasminogen binding lipoprotein of Bacteroides fragilis Chapter 7 Summarizing discussion 127 References 143 Nederlandse samenvatting 161 Dankwoord 167 Chapter 1 General introduction Chapter 1 Introduction The discovery of antibiotics was an enormous advance in medicine, which enabled mankind to fight bacterial diseases that could not be eradicated before. Nevertheless, with the ongoing process of bacterial resistance against (new) antibiotics, bacterial infections still pose a significant threat to human and animal health. Bacterial infections in humans can be categorized in multiple ways. A very easy way is the distinction between mono-infections, where only one bacterial species is the causative agent, and polymicrobial infections, where multiple bacterial species can be isolated from the site of infection. Mono-infections are well known to the general public because of news reports about Salmonella in chicken meat, methicillin resistant Staphylococcus aureus (MRSA) in hospitals and childhood immunization programs. Although less known to the public, community acquired as well as hospital acquired polymicrobial infections occur very frequently and can pose serious threats to human health. This thesis is focused on polymicrobial infections in the human intra- abdominal cavity. The abdomen is located between the thorax and the pelvis. The abdominal cavity is lined by the peritoneum: a serous sac, consisting of mesothelium and a thin layer of irregular connective tissue, that covers most of the viscera (internal organs) contained therein. Some abdominal organs, like the kidneys, are located retroperitoneally, but retroperitoneal infections are not considered here. Intra-abdominal infections and peritonitis In clinical practice, the term intra-abdominal infection (IAI) is often used as a synonym for peritonitis. However, these two are not the same (Fig. 1). Peritonitis is defined as an inflammation of the peritoneum, but this is not necessarily caused by bacteria (Farthmann and Schöffel, 1998). IAIs comprise a heterogeneous group of conditions encompassing bacterial peritonitis, intra-abdominal abscesses and infections of intra-abdominal organs like appendicitis and pancreatitis (Farthmann and Schöffel, 1998). IAIs are a common cause of hospitalization with approximately two million intra-abdominal procedures carried out annually in the USA (Onderdonk et al., 1990). These infections can be community acquired (e.g. appendicitis, stab wounds or traffic accidents) or hospital acquired, often as a complication of intra- abdominal surgery where the contents of the gut came into contact with the peritoneal cavity. 8 General introduction Fig. 1. Bacterial peritonitis in relation to intra-abdominal infections and peritonitis. The morbidity and mortality of IAI is significant. Data from hospitals in The Netherlands show that there are over 5300 new patients with serious forms of peritonitis each year (Bosscha et al., 1999). Extrapolated to the western world, this means that there must be more than half a million cases per year. Intra-abdominal abscesses occur in approximately 25% of patients treated for generalized peritonitis (Reijnen et al., 2003). Depending on the anatomic origin of the bacterial contamination, mortality ranges from 5 tot 50% (Brook, 1989; Farthmann and Schöffel, 1998). Complications like bacteremia and multiple-organ failure (MOF) frequently occur, increasing mortality rates up to 80% (Anaya and Nathens, 2003). IAIs have been found in 25% of the patients with MOF on a surgical intensive care unit (ICU) (Darling et al., 1988). The high morbidity of peritonitis indicates that the annual cost of treatment of these infections is significant. Bacterial peritonitis can be divided into three classes (Farthmann and Schöffel, 1998; Marshall, 2004). Primary peritonitis refers to spontaneous bacterial invasion of the peritoneal cavity. This mainly occurs in cirrhotic and immuno- compromised patients and the infections are characteristically aerobic and monomicrobial (Wittmann et al., 1996). Secondary peritonitis refers to peritoneal infections secondary to intra-abdominal lesions like perforation of the hollow viscus, bowel necrosis or penetrating infectious processes. Tertiary peritonitis is defined as peritonitis arising at least 48 hours after the apparently successful treatment of primary or secondary peritonitis. It is characterized by persistent or recurrent 9 Chapter 1 infections with organisms of low intrinsic virulence that are different from the organisms involved in primary or secondary peritonitis and it has a significant mortality. Host defense systems Once peritoneal contamination occurs, the body attempts to defend against this invasion with powerful mechanisms devised to deal with breaches in the integrity of the gut (Table 1) (reviewed by Hall et al., 1998). The first mechanism involves physical clearance of the infectious agent using the circa 100 ml of serous peritoneal fluid (Robinson, 1962). Bacteria are removed through gaps (stomata) in the mesothelial lining of the diaphragm into the lymph system (Tsilibary and Wissig, 1983; Nakatani et al., 1996). The second mechanism is destruction of the bacteria using the innate immune system. Complement activation is involved in the opsonization of microorganisms, enhancement of inflammatory responses, clearance of immune complexes and cell lysis (Pascual and French, 1995). The migration of polymorphonuclear neutrophils (PMNs) into the peritoneum is enhanced and also other cells of the innate immune system like macrophages and mast cells are involved (Caldwell and Watson, 1994). Table 1. Host defense systems in peritoneal infections System Mechanism 1. Clearance Serous peritoneal fluid Stomata 2. Innate immune system Polymorphonuclear neutrophyls Macrophages and mast cells 3. Abscess formation Coagulation favored over fibrinolysis Fibrin clot formation The third defense mechanism of the peritoneal cavity is containment of the infection as an abscess. This is the result of a shift in the balance between the coagulation
Load more

Recommended publications
  • Autodisplay: One-Component System for Efficient Surface Display And
    JOURNAL OF BACTERIOLOGY, Feb. 1997, p. 794–804 Vol. 179, No. 3 0021-9193/97/$04.0010 Copyright q 1997, American Society for Microbiology Autodisplay: One-Component System for Efficient Surface Display and Release of Soluble Recombinant Proteins from Escherichia coli 1 2 1,2 JOCHEN MAURER, JOACHIM JOSE, AND THOMAS F. MEYER * Downloaded from Abteilung Infektionsbiologie, Max-Planck-Institut fu¨r Biologie, 72076 Tu¨bingen,1 and Abteilung Molekulare Biologie, Max-Planck-Institut fu¨r Infektionsbiologie, 10117 Berlin,2 Germany Received 26 August 1996/Accepted 24 September 1996 The immunoglobulin A protease family of secreted proteins are derived from self-translocating polypro- tein precursors which contain C-terminal domains promoting the translocation of the N-terminally attached passenger domains across gram-negative bacterial outer membranes. Computer predictions identified the C-terminal domain of the Escherichia coli adhesin involved in diffuse adherence (AIDA-I) as a member of the autotransporter family. A model of the b-barrel structure, proposed to be responsible for http://jb.asm.org/ outer membrane translocation, served as a basis for the construction of fusion proteins containing heterologous passengers. Autotransporter-mediated surface display (autodisplay) was investigated for the cholera toxin B subunit and the peptide antigen tag PEYFK. Up to 5% of total cellular protein was detectable in the outer membrane as passenger autotransporter fusion protein synthesized under control of the constitutive PTK promoter. Efficient presentation of the passenger domains was demonstrated in the outer membrane protease T-deficient (ompT) strain E. coli UT5600 and the ompT dsbA double mutant JK321. Surface exposure was ascertained by enzyme-linked immunosorbent assay, immunofluorescence microscopy, and immunogold electron microscopy using antisera specific for the passenger domains.
    [Show full text]
  • Protein Secretion Systems in Fusobacterium Nucleatum
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Elsevier - Publisher Connector Biochimica et Biophysica Acta 1713 (2005) 92 – 112 http://www.elsevier.com/locate/bba Protein secretion systems in Fusobacterium nucleatum: Genomic identification of Type 4 piliation and complete Type V pathways brings new insight into mechanisms of pathogenesis Mickae¨l Desvauxa,b, Arshad Khana, Scott A. Beatsona, Anthony Scott-Tuckera, Ian R. Hendersona,* aThe Institute for Biomedical Research (IBR), The University of Birmingham-The Medical School, Division of Immunity and Infection, Bacterial Pathogenesis and Genomics Unit, Edgbaston, Birmingham B15 2TT, UK bInstitut National de la Recherche Agronomique (INRA), Centre de Recherches de Clermont-Ferrand-Theix, Unite´ de Recherche 370, Equipe Microbiologie, F-63122 Saint-Gene`s Champanelle, France Received 23 February 2005; received in revised form 11 April 2005; accepted 2 May 2005 Available online 8 June 2005 Abstract Recent genomic analyses of the two sequenced strains F. nucleatum subsp. nucleatum ATCC 25586 and F. nucleatum subsp. vincentii ATCC 49256 suggested that the major protein secretion systems were absent. However, such a paucity of protein secretion systems is incongruous with F. nucleatum pathogenesis. Moreover, the presence of one or more such systems has been described for every other Gram- negative organism sequenced to date. In this investigation, the question of protein secretion in F. nucleatum was revisited. In the current study, the absence in F. nucleatum of a twin-arginine translocation system (TC #2.A.64.), a Type III secretion system (TC #3.A.6.), a Type IV secretion system (TC #3.A.7.) and a chaperone/usher pathway (TC #1.B.11.) was confirmed.
    [Show full text]
  • Supplementary Information
    Supplementary information (a) (b) Figure S1. Resistant (a) and sensitive (b) gene scores plotted against subsystems involved in cell regulation. The small circles represent the individual hits and the large circles represent the mean of each subsystem. Each individual score signifies the mean of 12 trials – three biological and four technical. The p-value was calculated as a two-tailed t-test and significance was determined using the Benjamini-Hochberg procedure; false discovery rate was selected to be 0.1. Plots constructed using Pathway Tools, Omics Dashboard. Figure S2. Connectivity map displaying the predicted functional associations between the silver-resistant gene hits; disconnected gene hits not shown. The thicknesses of the lines indicate the degree of confidence prediction for the given interaction, based on fusion, co-occurrence, experimental and co-expression data. Figure produced using STRING (version 10.5) and a medium confidence score (approximate probability) of 0.4. Figure S3. Connectivity map displaying the predicted functional associations between the silver-sensitive gene hits; disconnected gene hits not shown. The thicknesses of the lines indicate the degree of confidence prediction for the given interaction, based on fusion, co-occurrence, experimental and co-expression data. Figure produced using STRING (version 10.5) and a medium confidence score (approximate probability) of 0.4. Figure S4. Metabolic overview of the pathways in Escherichia coli. The pathways involved in silver-resistance are coloured according to respective normalized score. Each individual score represents the mean of 12 trials – three biological and four technical. Amino acid – upward pointing triangle, carbohydrate – square, proteins – diamond, purines – vertical ellipse, cofactor – downward pointing triangle, tRNA – tee, and other – circle.
    [Show full text]
  • Conserved Features in the Structure, Mechanism, and Biogenesis of the Inverse Autotransporter Protein Family
    GBE Conserved Features in the Structure, Mechanism, and Biogenesis of the Inverse Autotransporter Protein Family Eva Heinz1,2,y, Christopher J. Stubenrauch1,y, Rhys Grinter1,3, Nathan P. Croft4, Anthony W. Purcell4, Richard A. Strugnell5, Gordon Dougan2, and Trevor Lithgow1,* 1Department of Microbiology, Infection & Immunity Program, Biomedicine Discovery Institute, Monash University, Clayton, Australia 2 Wellcome Trust Sanger Institute, Hinxton, United Kingdom Downloaded from https://academic.oup.com/gbe/article-abstract/8/6/1690/2574022 by guest on 13 December 2018 3Institute of Microbiology and Infection, School of Immunity and Infection, University of Birmingham, Birmingham, United Kingdom 4Department of Biochemistry and Molecular Biology, Infection & Immunity Program, Biomedicine Discovery Institute, Monash University, Clayton, Australia 5Department of Microbiology & Immunology, University of Melbourne, Parkville, Australia *Corresponding author: E-mail: [email protected]. yThese authors contributed equally to this work. Accepted: May 3, 2016 Abstract The bacterial cell surface proteins intimin and invasin are virulence factors that share a common domain structure and bind selectively to host cell receptors in the course of bacterial pathogenesis. The b-barrel domains of intimin and invasin show significant sequence and structural similarities. Conversely, a variety of proteins with sometimes limited sequence similarity have also been annotated as “intimin-like” and “invasin” in genome datasets, while other recent work on apparently unrelated virulence-associated proteins ultimately revealed similarities to intimin and invasin. Here we characterize the sequence and structural relationships across this complex protein family. Surprisingly, intimins and invasins represent a very small minority of the sequence diversity in what has been previously the “intimin/invasin protein family”.
    [Show full text]
  • Autodisplay of Enzymes—Molecular Basis and Perspectives
    Journal of Biotechnology 161 (2012) 92–103 Contents lists available at SciVerse ScienceDirect Journal of Biotechnology j ournal homepage: www.elsevier.com/locate/jbiotec Autodisplay of enzymes—Molecular basis and perspectives a,∗ b a Joachim Jose , Ruth Maria Maas , Mark George Teese a Institut für Pharmazeutische und Medizinische Chemie, Westfälische Wilhelms-Universität Münster, D-48149 Münster, Germany b Autodisplay Biotech GmbH, Merowingerplatz 1a, D-40225 Düsseldorf, Germany a r t i c l e i n f o a b s t r a c t Article history: To display an enzyme on the surface of a living cell is an important step forward towards a broader use of Received 8 October 2011 biocatalysts. Enzymes immobilized on surfaces appeared to be more stable compared to free molecules. It Received in revised form 14 February 2012 is possible by standard techniques to let the bacterial cell (e.g. Escherichia coli) decorate its surface with the Accepted 4 April 2012 enzyme and produce it on high amounts with a minimum of costs and equipment. Moreover, these cells Available online 30 April 2012 can be recovered and reused in several subsequent process cycles. Among other systems, autodisplay has some extra features that could overcome limitations in the industrial applications of enzymes. One major Keywords: advantage of autodisplay is the motility of the anchoring domain. Enzyme subunits exposed at the cell Autodisplay Biocatalysis surface having affinity to each other will spontaneously form dimers or multimers. Using autodisplay Synthesis enzymes with prosthetic groups can be displayed, expanding the application of surface display to the 5 6 Enzymes industrial important P450 enzymes.
    [Show full text]
  • Polarity and Secretion of Shigella Flexneri Icsa: a Classical Autotransporter
    Polarity and Secretion of Shigella flexneri IcsA: A Classical Autotransporter MATTHEW THOMAS DOYLE, B. SC. (BIOTECHNOLOGY) Submitted for the degree of Doctor of Philosophy Department of Molecular and Cellular Biology School of Biological Sciences The University of Adelaide Adelaide, South Australia, Australia July 2015 Declaration I certify that this Thesis contains no material which has been accepted for the award of any other degree or diploma in my name, in any university or other tertiary institution and, to the best of my knowledge and belief, contains no material previously published or written by another person, except where due reference has been made in the text. I certify that no part of this work will, in the future, be used in a submission in my name for any other degree or diploma in any university or other tertiary institution without the prior approval of the University of Adelaide and where applicable, any partner institution responsible for the joint- award of this degree. I give consent to this copy of my thesis when deposited in the University Library, being made available for loan and photocopying, subject to the provisions of the Copyright Act 1968. I acknowledge that copyright of published works contained within this thesis resides with the copyright holders of those works. I give permission for the digital version of my thesis to be made available on the web, via the University’s digital research repository, the Library Search and also through web search engines, unless permission has been granted by the University to restrict access for a period of time.
    [Show full text]
  • Folding Control in the Path of Type 5 Secretion
    toxins Review Folding Control in the Path of Type 5 Secretion Nathalie Dautin 1,2 1 Laboratoire de Biologie Physico-Chimique des Protéines Membranaires, Université de Paris, LBPC-PM, CNRS, UMR7099, 75005 Paris, France; [email protected] 2 Institut de Biologie Physico-Chimique, Fondation Edmond de Rothschild pour le Développement de la Recherche Scientifique, 75005 Paris, France Abstract: The type 5 secretion system (T5SS) is one of the more widespread secretion systems in Gram-negative bacteria. Proteins secreted by the T5SS are functionally diverse (toxins, adhesins, enzymes) and include numerous virulence factors. Mechanistically, the T5SS has long been considered the simplest of secretion systems, due to the paucity of proteins required for its functioning. Still, despite more than two decades of study, the exact process by which T5SS substrates attain their final destination and correct conformation is not totally deciphered. Moreover, the recent addition of new sub-families to the T5SS raises additional questions about this secretion mechanism. Central to the understanding of type 5 secretion is the question of protein folding, which needs to be carefully controlled in each of the bacterial cell compartments these proteins cross. Here, the biogenesis of proteins secreted by the Type 5 secretion system is discussed, with a focus on the various factors preventing or promoting protein folding during biogenesis. Keywords: secretion; folding; autotransporter; type 5 secretion system; intimin; invasin; two-partner secretion; trimeric autotransporter Key Contribution: This short review describes the different factors preventing or promoting folding of proteins secreted by the T5SS. Citation: Dautin, N. Folding Control in the Path of Type 5 Secretion.
    [Show full text]
  • The Autotransporter Secretion System Mickael Desvaux, Nicholas Parham, Ian Henderson
    The autotransporter secretion system Mickael Desvaux, Nicholas Parham, Ian Henderson To cite this version: Mickael Desvaux, Nicholas Parham, Ian Henderson. The autotransporter secretion system. Research in Microbiology, Elsevier, 2004, 155 (2), pp.53 - 60. 10.1016/j.resmic.2003.10.002. hal-02910767 HAL Id: hal-02910767 https://hal.inrae.fr/hal-02910767 Submitted on 3 Aug 2020 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. Research in Microbiology 155 (2004) 53–60 www.elsevier.com/locate/resmic Mini-review The autotransporter secretion system Mickaël Desvaux, Nicholas J. Parham, Ian R. Henderson ∗ Bacterial Pathogenesis and Genomics Unit, Division of Immunity and Infection, The Medical School, University of Birmingham, Edgbaston, Birmingham, B15 2TT, UK Received 9 September 2003; accepted 3 October 2003 First published online 7 October 2003 Abstract The type V secretion system includes the autotransporter family, the two-partner system and the Oca family. The autotransporter secretion process involving first the translocation of the precursor through the inner membrane and then its translocation through the outer membrane via a pore formed by a β-barrel is reviewed. 2003 Elsevier SAS. All rights reserved. Keywords: Autotransporter; Membrane transport protein; Protein translocation; Virulence factors 1.
    [Show full text]
  • Recent Advances in the Understanding of Trimeric Autotransporter Adhesins
    Medical Microbiology and Immunology (2020) 209:233–242 https://doi.org/10.1007/s00430-019-00652-3 REVIEW Recent advances in the understanding of trimeric autotransporter adhesins Andreas R. Kiessling1 · Anchal Malik1 · Adrian Goldman1,2 Received: 12 August 2019 / Accepted: 30 November 2019 / Published online: 21 December 2019 © The Author(s) 2019 Abstract Adhesion is the initial step in the infection process of gram-negative bacteria. It is usually followed by the formation of bioflms that serve as a hub for further spread of the infection. Type V secretion systems engage in this process by binding to components of the extracellular matrix, which is the frst step in the infection process. At the same time they provide protec- tion from the immune system by either binding components of the innate immune system or by establishing a physical layer against aggressors. Trimeric autotransporter adhesins (TAAs) are of particular interest in this family of proteins as they pos- sess a unique structural composition which arises from constraints during translocation. The sequence of individual domains can vary dramatically while the overall structure can be very similar to one another. This patchwork approach allows research- ers to draw conclusions of the underlying function of a specifc domain in a structure-based approach which underscores the importance of solving structures of yet uncharacterized TAAs and their individual domains to estimate the full extent of functions of the protein a priori. Here, we describe recent advances in understanding the translocation process of TAAs and give an overview of structural motifs that are unique to this class of proteins.
    [Show full text]
  • Reevaluating the Fusobacterium Virulence Factor Landscape ​ ​ 1* 1* 1 1 2 Ariana Umana ,​ Blake E
    bioRxiv preprint doi: https://doi.org/10.1101/534297; this version posted January 29, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. Reevaluating the Fusobacterium Virulence Factor Landscape ​ ​ 1* 1* 1 1 2 Ariana Umana ,​ Blake E. Sanders ,​ Chris C. Yoo ,​ Michael A. Casasanta ,​ Barath Udayasuryan ,​ Scott 2 ​ 1,# ​ ​ ​ ​ S. Verbridge ,​ Daniel J. Slade ​ ​ 1 Department​ of Biochemistry, Virginia Polytechnic Institute and State University, Blacksburg, VA, USA. 2 ​ Laboratory​ of Integrative Tumor Ecology, and Virginia Tech – Wake Forest School of Biomedical Engineering and Sciences, Blacksburg, VA, USA *These authors contributed equally # To whom correspondence should be addressed: Dr. Daniel J. Slade, Department of Biochemistry, Virginia Polytechnic Institute and ​ State University, Blacksburg, VA 24061. Telephone: +1 (540) 231-2842. Email: [email protected] ​ ________________________________________________________________________________________ ABSTRACT Fusobacterium are Gram-negative, anaerobic, opportunistic pathogens involved in ​ multiple diseases, including the oral pathogen Fusobacterium nucleatum being linked to the ​ progression and severity of colorectal cancer. The global identification of virulence factors in Fusobacterium has been greatly hindered by a lack of properly assembled and annotated genomes. Using newly completed genomes from nine strains and seven species of Fusobacterium, we report the ​ ​ identification and correction of virulence factors from the Type 5 secreted autotransporter and FadA protein families, with a focus on the genetically tractable strain F. nucleatum subsp. nucleatum ATCC ​ ​ ​ 23726 and the classic typed strain F. nucleatum subsp.
    [Show full text]
  • Structural and Functional Analysis of Moraxella Catarrhalis Adhesins MCAP and OMPCD
    Health Science Campus FINAL APPROVAL OF DISSERTATION Doctor of Philosophy in Biomedical Sciences Structural and Functional Analysis of Moraxella catarrhalis Adhesins MCAP and OMPCD Submitted by: Chritine Akimana In partial fulfillment of the requirements for the degree of Doctor of Philosophy in Biomedical Sciences Examination Committee Major Advisor: Eric Lafontaine, Ph.D. Academic Robert Blumenthal, Ph.D. Advisory Committee: R. Mark Wooten, Ph.D. Ray Larsen, Ph.D. Venkatesha Basrur, Ph.D. Senior Associate Dean College of Graduate Studies Michael S. Bisesi, Ph.D. Date of Defense: April 16, 2007 Structural and Functional Analysis of Moraxella catarrhalis Adhesins McaP and OMPCD Christine Akimana University of Toledo Health Science Campus 2007 DEDICATION This dissertation is dedicated to my son, Bennett Kiza, who has filled my life with untold blessings. I would like to thank my husband, Nicolas Kiza, and my mother-in-law, whose support and encouragement has enabled me to complete this work. I also would like to thank my parents for their love, inspiration, and support, especially for their hard work that has enabled me to achieve so much in life. There are several people, family members, friends, and mentors, whose support and inspiration has helped me throughout my academic years. For those as well, I offer my full gratitude. ii ACKNOWLEDGEMENTS I would like to thank my major advisor Dr. Eric Lafontaine for giving me the opportunity to work on this very exciting project. I thank him for training me, for sharing his expertise, and for providing me with the opportunity to attend several scientific conferences. I also thank him for his kindness, support, and patience.
    [Show full text]
  • Two-Partner Secretion: Combining Efficiency and Simplicity in the Secretion of Large Proteins for Bacteria-Host and Bacteria-Bacteria Interactions
    REVIEW published: 09 May 2017 doi: 10.3389/fcimb.2017.00148 Two-Partner Secretion: Combining Efficiency and Simplicity in the Secretion of Large Proteins for Bacteria-Host and Bacteria-Bacteria Interactions Jeremy Guérin 1, Sarah Bigot 2, Robert Schneider 3, Susan K. Buchanan 1 and Françoise Jacob-Dubuisson 4* 1 Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, USA, 2 Molecular Microbiology and Structural Biochemistry, Centre National de La Recherche Scientifique UMR 5086–Université Lyon 1, Institute of Biology and Chemistry of Proteins, Lyon, France, 3 NMR and Molecular Interactions, Université de Lille, Centre National de La Recherche Scientifique, UMR 8576–Unité de Glycobiologie Structurale et Fonctionnelle, Lille, France, 4 Université de Lille, Centre National de La Recherche Scientifique, Institut National de La Santé et de La Recherche Médicale, CHU Lille, Institut Pasteur de Lille, U1019–UMR 8204–Centre d’Infection et d’Immunité de Lille, Lille, France Edited by: Sophie Bleves, Initially identified in pathogenic Gram-negative bacteria, the two-partner secretion (TPS) Aix-Marseille University, France pathway, also known as Type Vb secretion, mediates the translocation across the outer Reviewed by: membrane of large effector proteins involved in interactions between these pathogens Raffaele Ieva, UMR5100 Laboratoire de and their hosts. More recently, distinct TPS systems have been shown to secrete toxic Microbiologie et Génétique effector domains that participate in inter-bacterial competition or cooperation. The effects Moléculaires, France of these systems are based on kin vs. non-kin molecular recognition mediated by Dirk Linke, University of Oslo, Norway specific immunity proteins.
    [Show full text]