The Hns Gene of Escherichia Coli Is Transcriptionally Down-Regulated by (P)Ppgpp
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Assessing the Role of Spot and Rela in Capsular Polysaccharide
Journal of Experimental Microbiology and Immunology (JEMI) Vol. 15: 52 – 58 Copyright © April 2011, M&I UBC Assessing the Role of SpoT and RelA in Capsular Polysaccharide Synthesis after Treatment with Sub-lethal Concentrations of Kanamycin to Confer Decreased Antibiotic Sensitivity in Escherichia coli Wesley Chenne, Louisa Ng, and Mary Rose Pambid Department of Microbiology & Immunology, University of British Columbia Resistance to various antibiotics has been associated with increased capsular polysaccharide production through activation of RelA and SpoT, regulators of the stringent response, through their respective synthesis and hydrolysis of guanosine tetraphosphate in Escherichia coli. In order to further characterize the role of SpoT and RelA in capsular polysaccharide production and in conferring antibiotic resistance, growth patterns and induced capsular polysaccharide levels of various strains were determined following sub- lethal treatment with kanamycin and further treatment at inhibitory levels of kanamycin. Contrary to previous reports that capsular polysaccharide confers antibiotic resistance, it was found that spoT mutants produced the least capsular polysaccharide but had higher levels of resistance in comparison to relA mutants, which produced the most polysaccharide but exhibited lower resistance. As expected, both these mutants exhibited lower resistance to kanamycin as a result of pre-treatment than the wild-type strain. Thus, we propose that bacterial survival and resistance development upon antibiotic administration -
Développement D'hybrides Aptamère-Peptide
UNIVERSITÉ DU QUÉBEC INSTITUT NATIONAL DE LA RECHERCHE SCIENTIFIQUE INSTITUT ARMAND-FRAPPIER DÉVELOPPEMENT D’HYBRIDES APTAMÈRE-PEPTIDE ANTIMICROBIEN UN MODÈLE POUR CIBLER DES BACTÉRIES PATHOGÈNES Par Amal Thamri Mémoire présenté pour l’obtention du grade de Maître en sciences (M.Sc.) en microbiologie appliquée Jury d’évaluation Examinateur externe: Dr. Roger.C Lévesque, Université de Laval Examinateur interne: Dr. Frédéric Veyrier, INRS-IAF Directeur de recherche: Dr. Jonathan Perreault, INRS-IAF Co-directrice de recherche: Dre. Annie Castonguay, INRS-IAF Remerciements Cette maîtrise a été réalisée dans le cadre d’une coopération entre INRS et le ministère d’enseignement supérieur tunisien. Les recherches objets de ce mémoire ont été réalisées dans différents laboratoires de l’institut Armand-Frappier: Laboratoires des professeurs: Jonathan Perreault, Annie Castonguay, Eric Déziel, David Chatenet. Je tiens à remercier toutes les personnes qui ont participé de près ou de loin au bon déroulement et à l’avancement des travaux de cette maîtrise. Je remercie spécialement mon directeur de recherche Jonathan Perreault pour avoir cru en mes capacités d’intégrer sa jeune équipe et travailler sur un projet novateur. Je remercie très sincèrement ma co-directrice Annie Castonguay pour ses encouragements tout au long de ces deux années. Je les remercie également d'avoir bien assuré la direction et l'encadrement de mes travaux de recherche. Merci au professeur Eric Déziel pour sa compréhension et ses conseils précieux. J’adresse aussi mes remerciements au professeur David Chatenet pour sa disponibilité, ses directions bénéfiques et ses remarques pertinentes. J’adresse mes sentiments de respect et de gratitude à Myriam Letourneau et Marie- Christine Groleau pour leur générosité et leur aide à mettre en place et améliorer plusieurs expériences. -
The Alarmone (P)Ppgpp Is Part of the Heat Shock Response of Bacillus
bioRxiv preprint doi: https://doi.org/10.1101/688689; this version posted July 1, 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 4.0 International license. 1 The alarmone (p)ppGpp is part of the heat shock response of Bacillus 2 subtilis 3 4 Heinrich Schäfer1,2, Bertrand Beckert3, Wieland Steinchen4, Aaron Nuss5, Michael 5 Beckstette5, Ingo Hantke1, Petra Sudzinová6, Libor Krásný6, Volkhard Kaever7, Petra 6 Dersch5,8, Gert Bange4*, Daniel Wilson3* & Kürşad Turgay1,2* 7 8 Author affiliations: 9 1 Institute of Microbiology, Leibniz Universität Hannover, Herrenhäuser Str. 2, D-30419, 10 Hannover, Germany. 11 2 Max Planck Unit for the Science of Pathogens, Charitéplatz 1, 10117 Berlin, Germany 12 3 Institute for Biochemistry and Molecular Biology, Martin-Luther-King Platz 6, University of 13 Hamburg, 20146 Hamburg, Germany. 14 4 Philipps-University Marburg, Center for Synthetic Microbiology (SYNMIKRO) and 15 Department of Chemistry, Hans-Meerwein-Strasse, 35043 Marburg, Germany. 16 5 Department of Molecular Infection Biology, Helmholtz Centre for Infection Research, 17 Braunschweig, Germany 18 6 Institute of Microbiology, Czech Academy of Sciences, Vídeňská 1083, 142 20, Prague, 19 Czech Republic. 20 7 Hannover Medical School, Research Core Unit Metabolomics, Carl-Neuberg-Strasse 1, 21 30625, Hannover, Germany. 22 8 Institute of Infectiology, Von-Esmarch-Straße 56, University of Münster, Münster, Germany 23 24 Short title: The interplay of heat shock and stringent response 25 1 bioRxiv preprint doi: https://doi.org/10.1101/688689; this version posted July 1, 2019. -
(P)Ppgpp Synthesis by the Ca2+-Dependent Rela-Spot Homolog
bioRxiv preprint doi: https://doi.org/10.1101/767004; this version posted September 12, 2019. 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. Plastidial (p)ppGpp synthesis by the Ca2+-dependent RelA-SpoT homolog regulates the adaptation of chloroplast gene expression to darkness in Arabidopsis Sumire Ono1,4, Sae Suzuki1,4, Doshun Ito1 Shota Tagawa2, Takashi Shiina2, and Shinji Masuda3,5 1School of Life Science & Technology, Tokyo Institute of Technology, Yokohama 226-8501, Japan 2Graduate School of Life and Environmental Sciences, Kyoto Prefectural University, Sakyo-ku, Kyoto 606-8522, Japan 3Center for Biological Resources & Informatics, Tokyo Institute of Technology, Yokohama 226-8501, Japan 4These authors contributed equally to the study 5Corresponding author: E-mail, [email protected] Abbreviations: CRSH, Ca2+-activated RSH; flg22, flagellin 22; (p)ppGpp, 5'-di(tri)phosphate 3'-diphosphate; PAMPs; pathogen-associated molecular patterns; RSH, RelA-SpoT homolog; qRT-PCR, quantitative real-time PCR 1 bioRxiv preprint doi: https://doi.org/10.1101/767004; this version posted September 12, 2019. 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. Abstract In bacteria, the hyper-phosphorylated nucleotides, guanosine 5'-diphosphate 3'-diphosphate (ppGpp) and guanosine 5'-triphosphate 3'-diphosphate (pppGpp), function as secondary messengers in the regulation of various metabolic processes of the cell, including transcription, translation, and enzymatic activities, especially under nutrient deficiency. The activity carried out by these nucleotide messengers is known as the stringent response. -
The Link Between Purine Metabolism and Production of Antibiotics in Streptomyces
antibiotics Review The Link between Purine Metabolism and Production of Antibiotics in Streptomyces Smitha Sivapragasam and Anne Grove * Department of Biological Sciences, Louisiana State University, Baton Rouge, LA 70803, USA; [email protected] * Correspondence: [email protected] Received: 10 May 2019; Accepted: 3 June 2019; Published: 6 June 2019 Abstract: Stress and starvation causes bacterial cells to activate the stringent response. This results in down-regulation of energy-requiring processes related to growth, as well as an upregulation of genes associated with survival and stress responses. Guanosine tetra- and pentaphosphates (collectively referred to as (p)ppGpp) are critical for this process. In Gram-positive bacteria, a main function of (p)ppGpp is to limit cellular levels of GTP, one consequence of which is reduced transcription of genes that require GTP as the initiating nucleotide, such as rRNA genes. In Streptomycetes, the stringent response is also linked to complex morphological differentiation and to production of secondary metabolites, including antibiotics. These processes are also influenced by the second messenger c-di-GMP. Since GTP is a substrate for both (p)ppGpp and c-di-GMP, a finely tuned regulation of cellular GTP levels is required to ensure adequate synthesis of these guanosine derivatives. Here, we discuss mechanisms that operate to control guanosine metabolism and how they impinge on the production of antibiotics in Streptomyces species. Keywords: c-di-GMP; guanosine and (p)ppGpp; purine salvage; secondary metabolism; Streptomycetes; stringent response 1. Introduction Bacteria experience constant challenges, either in the environment or when infecting a host. They utilize various mechanisms to survive such stresses, which may include changes in temperature, pH, or oxygen content as well as limited access to carbon or nitrogen sources. -
Quantification of Guanosine Triphosphate and Tetraphosphate In
Quantification of guanosine triphosphate and tetraphosphate in plants and algae using stable isotope-labelled internal standards Julia Bartoli, Sylvie Citerne, Gregory Mouille, Emmanuelle Bouveret, Ben Field To cite this version: Julia Bartoli, Sylvie Citerne, Gregory Mouille, Emmanuelle Bouveret, Ben Field. Quantification of guanosine triphosphate and tetraphosphate in plants and algae using stable isotope-labelled internal standards. Talanta, Elsevier, 2020, 219, pp.121261. 10.1016/j.talanta.2020.121261. cea-02961710 HAL Id: cea-02961710 https://hal-cea.archives-ouvertes.fr/cea-02961710 Submitted on 19 Nov 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. Quantification of guanosine triphosphate and tetraphosphate in plants and algae using stable isotope- labelled internal standards. Julia Bartoli1, Sylvie Citerne2, Gregory Mouille2, Emmanuelle Bouveret3 and Ben Field4* 1Aix Marseille Univ CNRS, LISM, UMR 7255, IMM FR 3479, 31 Chemin Joseph Aiguier, 13009, Marseille, France 2 Institut Jean-Pierre Bourgin, INRAE, AgroParisTech, Université Paris-Saclay, -
Guanosine Pentaphosphate Phosphohydrolase of Escherichia Coli Is a Long-Chain Exopolyphosphatase J
Proc. Natl. Acad. Sci. USA Vol. 90, pp. 7029-7033, August 1993 Biochemistry Guanosine pentaphosphate phosphohydrolase of Escherichia coli is a long-chain exopolyphosphatase J. D. KEASLING*, LEROY BERTSCHt, AND ARTHUR KORNBERGtI *Department of Chemical Engineering, University of California, Berkeley, CA 94720-9989; and tDepartment of Biochemistry, Stanford University School of Medicine, Stanford, CA 94305-5307 Contributed by Arthur Kornberg, April 14, 1993 ABSTRACT An exopolyphosphatase [exopoly(P)ase; EC MATERIALS AND METHODS 3.6.1.11] activity has recently been purified to homogeneity from a mutant strain of Escherichia coi which lacks the Reagents and Proteins. Sources were as follows: ATP, principal exopoly(P)ase. The second exopoly(P)ase has now ADP, nonradiolabeled nucleotides, poly(P)s, bovine serum been identified as guanosine pentaphosphate phosphohydro- albumin, and ovalbumin from Sigma; [y-32P]ATP at 6000 lase (GPP; EC 3.6.1.40) by three lines of evidence: (i) the Ci/mmol (1 Ci = 37 GBq) and [y-32P]GTP at 6000 Ci/mmol sequences of five btptic digestion fragments of the purified from ICN; Q-Sepharose fast flow, catalase, aldolase, Super- protein are found in the translated gppA gene, (u) the size ofthe ose-12 fast protein liquid chromatography (FPLC) column, protein (100 kDa) agrees with published values for GPP, and and Chromatofocusing column and reagents from Pharmacia (iu) the ratio of exopoly(P)ase activity to GPP activity remains LKB; DEAE-Fractogel, Pll phosphocellulose, and DE52 constant throughout a 300-fold purification in the last steps of DEAE-cellulose from Whatman; protein standards for SDS/ the procedure. -
N. Meningitidis
Stringent response regulation and its impact on ex vivo survival in the commensal pathogen Neisseria meningitidis Regulation der stringenten Kontrolle und ihre Auswirkungen auf das ex vivo Überleben des kommensalen Erregers Neisseria meningitidis Dissertation zur Erlangung des naturwissenschaftlichen Doktorgrades der Julius-Maximilians-Universität Würzburg vorgelegt von Laura Violetta Hagmann (geb. Kischkies) Frankfurt am Main Würzburg, 2016 Eingereicht am: …………………………… Mitglieder der Promotionskommission: Vorsitzender: …………………………………………………….. Gutachter: PD Dr. rer. nat. Dr. med. Christoph U. Schoen Gutachter: PD Dr. rer. nat. Knut Ohlsen Tag des Promotionskolloquiums: …………………………... Doktorurkunde ausgehändigt am: …………………………... EIDESSTATTLICHE ERKLÄRUNG Hiermit versichere ich, dass ich die vorliegende Dissertation selbstständig angefertigt und nur die angegebenen Quellen und Hilfsmittel verwendet habe. Ich versichere zudem, dass diese Arbeit in dieser oder ähnlicher Form in keinem anderen Prüfungsverfahren vorgelegen hat. Bis auf den Titel der Diplom-Biologin habe ich bislang keinen anderen akademischen Grad erworben oder zu erwerben versucht. Würzburg, den 13.10.2016 ……………………………………. Laura Violetta Hagmann Table of Content 1 Summary .......................................................................................................... 1 2 Zusammenfassung .......................................................................................... 2 3 Introduction..................................................................................................... -
Ribosomal RNA
Ribosomal RNA Ribosomal ribonucleic acid (rRNA) is a type of non-coding RNA which is the primary component of ribosomes, essential to all cells. rRNA is a ribozyme which carries out protein synthesis in ribosomes. Ribosomal RNA is transcribed from ribosomal DNA (rDNA) and then bound to ribosomal proteins to form small and large ribosome subunits. rRNA is the physical and mechanical factor of the ribosome that forces transfer RNA (tRNA) and messenger RNA (mRNA) to process and translate the latter into proteins.[1] Ribosomal RNA Three-dimensional views of the ribosome, showing rRNA in dark blue (small subunit) is the predominant form of RNA found in most cells; it makes and dark red (large subunit). Lighter colors up about 80% of cellular RNA despite never being translated represent ribosomal proteins. into proteins itself. Ribosomes are composed of approximately 60% rRNA and 40% ribosomal proteins by mass. Contents Structure Assembly Function Subunits and associated ribosomal RNA In prokaryotes In eukaryotes Biosynthesis In eukaryotes Eukaryotic regulation In prokaryotes Prokaryotic regulation Degradation In eukaryotes In prokaryotes Sequence conservation and stability Significance Human genes See also References External links Structure Although the primary structure of rRNA sequences can vary across organisms, base-pairing within these sequences commonly forms stem-loop configurations. The length and position of these rRNA stem-loops allow them to create three-dimensional rRNA structures that are similar across species.[2] Because of these configurations, rRNA can form tight and specific interactions with ribosomal proteins to form ribosomal subunits. These ribosomal proteins contain basic residues (as opposed to acidic residues) and aromatic residues (i.e. -
334429671.Pdf
View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by DSpace at Tartu University Library Talanta 205 (2019) 120161 Contents lists available at ScienceDirect Talanta journal homepage: www.elsevier.com/locate/talanta Analysis of nucleotide pools in bacteria using HPLC-MS in HILIC mode T Eva Zborníkováa,b, Zdeněk Knejzlíka, Vasili Hauryliukc,d,e, Libor Krásnýf, Dominik Rejmana,* a Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, v.v.i., Flemingovonam. 2, CZ-166 10, Prague 6, Czech Republic b Charles University, Faculty of Science, Department of Analytical Chemistry, Hlavova 8, 128 43, Prague 2, Czech Republic c Department of Molecular Biology, Umeå University, Building 6K, 6L University Hospital Area, SE-901 87, Umeå, Sweden d Laboratory for Molecular Infection Medicine Sweden (MIMS), Umeå University, Building 6K and 6L, University Hospital Area, 90187, Umeå, Sweden e University of Tartu, Institute of Technology, 50411, Tartu, Estonia f Institute of Microbiology, Czech Academy of Sciences v.v.i., Vídeňská 1083, 142 20, Prague 4, Czech Republic ARTICLE INFO ABSTRACT Keywords: Nucleotides, nucleosides and their derivatives are present in all cells at varying concentrations that change with Nucleotide the nutritional, and energetic status of the cell. Precise measurement of the concentrations of these molecules is HPLC-MS instrumental for understanding their regulatory effects. Such measurement is challenging due to the inherent HILIC instability of these molecules and, despite many decades of research, the reported values differ widely. Here, we ppGpp present a comprehensive and easy-to-use approach for determination of the intracellular concentrations of > 25 Stringent response target molecular species. -
(P)Ppgpp in Plants and Algae Ben Field
Green magic: regulation of the chloroplast stress response by (p)ppGpp in plants and algae Ben Field To cite this version: Ben Field. Green magic: regulation of the chloroplast stress response by (p)ppGpp in plants and algae. Journal of Experimental Botany, Oxford University Press (OUP), 2018, 69 (11), pp.2797-2807. 10.1093/jxb/erx485. hal-01855838 HAL Id: hal-01855838 https://hal-amu.archives-ouvertes.fr/hal-01855838 Submitted on 8 Aug 2018 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. Green magic: regulation of the chloroplast stress response by (p)ppGpp in plants and algae Ben Field Aix Marseille Univ, CEA, CNRS, France Correspondence: [email protected] Abstract The hyperphosphorylated nucleotides guanosine pentaphosphate and tetraphosphate [together referred to as (p)ppGpp, or ‘magic spot’] orchestrate a signalling cascade in bacteria that controls growth under optimal conditions and in response to environmental stress. (p)ppGpp is also found in the chloroplasts of plants and algae where it has also been shown to accumulate in response to abiotic stress. Recent studies suggest that (p)ppGpp is a potent inhibitor of chloroplast gene expression in vivo, and is a significant regulator of chloroplast function that can influence both the growth and the development of plants. -
Role of the Stringent Response in Crop Plants Growth and Development
Tytuł projektu Rola odpowiedzi ścisłej w wzroście i rozwoju roślin uprawnych Project title Role of the stringent response in crop plants growth and development Dyscyplina /Area of science Nauki biologiczne/Biological Sciences PROJECT DESCRIPTION Project goals To find out which developmental stages during plant growth are affected/regulated by the stringent response To analyze whether changes in (p)ppGpp content always correlate with so far known (p)ppGpp-mediated changes in chloroplast functioning To introduce the fluorescence in situ hybridization method for the purpose of the stringent response analysis To find out plant growth stages that can be potentially regulated by means of agents controlling (p)ppGpp content Outline The stringent response was found to take place in chloroplasts less than 20 years ago. The effectors of the response are guanosine tetraphosphate and guanosine pentaphosphate (ppGpp and pppGpp), unusual nucleotides jointly referred to as (p)ppGpp or magic spots. These effectors are synthesized by nucleus-encoded RSH proteins that are known to function in chloroplasts. The research work of the last four years has shown that (p)ppGpp inhibit chloroplast transcription (e.g. of rRNA, tRNA, genes encoding proteins involved in translation and photosynthesis), translation and the production of many metabolites. The molecular changes invoked upon the accumulation of (p)ppGpp eventually cause a decrease in the efficiency of photosynthesis and the reduction of plant growth. However, still, the stages of plant development that are being affected with/regulated by those nucleotides are not known (reviewed in Boniecka et al., 2017, Planta 246:817-842). The aim of the project is to elucidate the role of the stringent response in plant development, in particular of rapeseed (Brassica napus L.), as it is an important crop plant belonging to the same family as the model plant Arabidopsis thaliana, and its genome has been sequenced.