DOCSLIB.ORG

Caracterización De La Agrupación Génica De Biosíntesis Del Inmunosupresor Tacrolimus En Dos Especies Del Género Streptomyces

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Caracterización De La Agrupación Génica De Biosíntesis Del Inmunosupresor Tacrolimus En Dos Especies Del Género Streptomyces Departamento de Biología Molecular Area de Microbiología Caracterización de la agrupación génica de biosíntesis del inmunosupresor tacrolimus en dos especies del género Streptomyces. Estudio de su regulación. TESIS DOCTORAL Elena Solera Segura León, 2014 Agradecimientos El desarrollo de esta tesis doctoral no hubiera sido posible sin el apoyo personal, técnico y académico de un gran número de personas que han facilitado de manera inestimable la realización de este trabajo. En primer lugar agradecer al Dr. Juan Francisco Martín, director de esta tesis, por creer en mí ya desde mi época de estudiante, por permitirme desarrollar este proyecto bajo su tutela. A Alberto Sola, codirector de esta Tesis, por introducirme y guiarme en el mundo de la investigación, por mostrarme su brillantez, por su supervisión tolerante con margén para la experimentación bajo iniciativa propia y por su meticulosa corrección de la memoria. Al Ministerio de Educación y Ciencia por la concesión de una beca predoctoral asociada al proyecto Bio2006-14853-c02-01, en el cual se enmarca este trabajo. A Patricia por su gran eficacia y eficiencia en la secuenciación de los cósmidos. Por estar siempre con una sonrisa en la cara y por ser una gran AMIGA. A Maite por su ayuda con las extracciones de ARN, por ser de esas amigas que sabes que nunca te fallan, sin duda lo mejor de la recta final de la Tesis. A Claudia por su ayuda en el análisis de las qRT-PCR, por ser una gran compañera de viaje y por estar siempre dispuesta a ayudar. A Javi por su ayuda con cualquier duda de laboratorio, por las charlas infinitas a última hora y por su estima. A Rubén por los buenos momentos compartidos. A Angelina por su calided y amistad. A Lara por ayudarme en los comienzos, aunque separadas los lazos siguen ahí. A Silvia por su siempre disposición y por su asistencia técnica con el HPLC. A Miriam y Zahara por su ayuda con el cultivo de Streptomyces tacrolimicus, hicimos lo que pudimos con nuestra querida cepa. A Carlos Barreiro por contar conmigo para participar en el proyecto de secuenciación y demás colaboraciones. A Antonio por su siempre paciencia y disposición. A Rosma por su claridad y buena conversación. Al resto de compañeros de INBIOTEC que habéis ayudado a crear un inmejorable ambiente de trabajo especialmente a Clara, Mar, Maria, Nuria y Tamara por vuestras amenas charlas. A Andrea, Bea, Alcira, Berna y Josefina por su profesionalidad y apoyo administrativo y técnico. A mis padres, mi referencia de esfuerzo y trabajo duro, gracias por proporcionarme un ambiente estable en el que poder desarrollarme, gracias por vuestro apoyo incondicional, soy lo que soy gracias a vosotros. A Rubenin, un claro ejemplo de que los límites no vienen impuestos sino que los marcamos nosotros mismos, eres nuestro orgullo. A Rubén y Vladi gracias por vuestro cariño. A la pequeña Sofía gracias por inundar la familia de alegría. A mis padres y hermanos Abreviaturas y acrónimos Se presentan a continuación las abreviaturas y acrónimos empleados a lo largo de esta memoria. Se excluyen en general las soluciones y medios descritos en el capítulo “Materiales y métodos”. AcNa: Acetato sódico HK: histidina quinasa ACP: proteína trasportadora de grupos HPLC: Cromatografía líquida de alta acilo resolución ADNc: ADN complementario HTH: hélice-vuelta-hélice ADNg: ADN total o genómico IPTG: 1-isopropil-β-D-galactopiranósido antibióticoR: resistente a un determinado kb: kilopares de bases antibiótico KCTC: colección de cultivos tipo koreana antibióticoS: sensible a un determinado Kn: kanamicina antibiótico KR: cetoreductasa Apr: apramicina KS: cetosintasa ARNr: ARN ribosomal LAL: proteína de gran tamaño de unión a ARNt: ARN de transferencia ATP de la familia LuxR AT: aciltransferasa M: molar. ATCC: colección americana de cultivos tipo MCS: sitio de clonación múltiple AU: unidades de absorbancia MMF: micofenolato de mofetilo AZA: azatioprina MTBE: metilterbutileter CIA: cloroformo-alcohol isoamílico nm: nanómetro CoA: coenzima A NRPS: péptido sintasa no ribosomal CsA: ciclosporina A Nt: N-terminal Ct: C-terminal nt: nucleótido CTAB: bromuro de cetil-trimetil amonio ORF: marco de lectura abierto DH: deshidratasa pb: pares de bases DCDC: 4,5 dihidroxi-1,5-dienocarboxílico PKS(s): policétido sintasa(s) DHCHC: ácido 4,5 dihidroxiciclohex-1- PT: proteínas terminales enecarboxílico qRT-PCR: PCR cuantitativa a tiempo real DMSO: dimetilsulfóxido QS: quórum sensing dNTPs: desoxinucleótidos trifosfato RIN: número de integridad del ARN EDTA: ácido etilendiaminotetraacético rpm: revoluciones por minuto ER: enoil reductasa RR: Regulador de respuesta FAS: sintasa de ácidos grasos SDS: docedil sulfato sódico FDA: agencia del medicamento de los EEUU Tª: Temperatura GlcNAc: N-acetil glucosamina TE: tioesteresa HK: Histidina quinasa TFA: ácido trifluoro acético TFR: familia reguladores transcripcionales TetR vol / v: volumen TIR: Repeticiones terminales invertidas WT: fenotipo salvaje (wild type). Tm: temperatura de fusión X-Gal: 5-bromo-4-cloro-3-indolil- Tris: tris-hidroximetil-aminometano -β-D-galactopiranósido Glosario de términos coloquiales en medicina pero que no están recogidos en la RAE: Sintenia Clado Índice Índice I 1. INTRODUCCIÓN ................................................................................................................................................. 1 1.1. GÉNERO STREPTOMYCES ................................................................................................................................... 3 1.1.1. TAXONOMÍA DEL GÉNERO ....................................................................................................................................... 3 1.1.2. CARACTERÍSTICAS BIOLÓGICAS DEL GÉNERO STREPTOMYCES ........................................................................................... 4 1.1.3. ECOLOGÍA DEL GÉNERO STREPTOMYCES ..................................................................................................................... 6 1.1.3.1. Importancia ecológica de Streptomyces ....................................................................................................... 7 1.1.4. CICLO DE VIDA DEL GÉNERO STREPTOMYCES ............................................................................................................... 8 1.1.5. CARACTERÍSTICAS GENÉTICAS DEL GÉNERO STREPTOMYCES .......................................................................................... 13 1.1.5.1. El cromosoma ............................................................................................................................................. 13 1.1.5.2. Estructura cromosómica ............................................................................................................................. 13 1.1.5.3. Organización del cromosoma ..................................................................................................................... 15 1.1.5.4. Cromosoma lineal e inestabilidad génica: ventaja adaptativa ................................................................... 16 1.1.5.5. Elementos genéticos extracromosomales .................................................................................................. 17 1.2. FÁRMACOS INMUNOSUPRESORES: TACROLIMUS............................................................................................ 19 1.2.1. HISTORIA DE LOS INMUNOSUPRESORES .................................................................................................................... 19 1.2.2. INDUSTRIA FARMACÉUTICA: IMPORTANCIA ECONÓMICA DE TACROLIMUS........................................................................ 23 1.2.3. BIOSÍNTESIS DE TACROLIMUS ................................................................................................................................. 25 1.2.3.1. Policétido sintasa de tacrolimus: síntesis de pre-tacrolimus ...................................................................... 25 1.2.3.2. Modificaciones post-PKS: Funcionalización de tacrolimus ......................................................................... 27 1.2.3.3. Tacrolimus y sus análogos .......................................................................................................................... 27 1.2.3.4. Rutas metabólicas implicadas en la biosíntesis de tacrolimus ................................................................... 28 1.2.4. ESPECIES PRODUCTORAS DE TACROLIMUS ................................................................................................................. 33 1.2.4.1. Streptomyces tsukubaensis ......................................................................................................................... 33 1.2.4.2. Streptomyces tacrolimicus .......................................................................................................................... 34 1.3. METABOLISMO SECUNDARIO Y SU REGULACIÓN ............................................................................................ 36 1.3.1. GÉNERO STREPTOMYCES: GRAN PRODUCTOR DE METABOLITOS SECUNDARIOS ................................................................ 36 1.3.2. REGULACIÓN DEL METABOLISMO SECUNDARIO EN STREPTOMYCES ................................................................................ 38 1.3.2.1. Reguladores globales .................................................................................................................................. 39 1.3.2.1.1. Reguladores globales de la diferenciación morfológica y la producción
Load more

Recommended publications
  • 341388717-Oa
    ORIGINAL RESEARCH published: 16 May 2017 doi: 10.3389/fmicb.2017.00877 Streptomyces colonosanans sp. nov., A Novel Actinobacterium Isolated from Malaysia Mangrove Soil Exhibiting Antioxidative Activity and Cytotoxic Potential against Human Colon Cancer Cell Lines Jodi Woan-Fei Law 1, Hooi-Leng Ser 1, Acharaporn Duangjai 2, 3, Surasak Saokaew 1, 3, 4, Sarah I. Bukhari 5, Tahir M. Khan 1, 6, Nurul-Syakima Ab Mutalib 7, Kok-Gan Chan 8, Edited by: Bey-Hing Goh 1, 3* and Learn-Han Lee 1, 3* Dongsheng Zhou, Beijing Institute of Microbiology and 1 Novel Bacteria and Drug Discovery Research Group, School of Pharmacy, Monash University Malaysia, Bandar Sunway, Epidemiology, China Malaysia, 2 Division of Physiology, School of Medical Sciences, University of Phayao, Phayao, Thailand, 3 Center of Health Reviewed by: Outcomes Research and Therapeutic Safety, School of Pharmaceutical Sciences, University of Phayao, Phayao, Thailand, 4 Andrei A. Zimin, Faculty of Pharmaceutical Sciences, Pharmaceutical Outcomes Research Center, Naresuan University, Phitsanulok, 5 6 Institute of Biochemistry and Thailand, Department of Pharmaceutics, College of Pharmacy, King Saud University, Riyadh, Saudi Arabia, Department of 7 Physiology of Microorganisms (RAS), Pharmacy, Absyn University Peshawar, Peshawar, Pakistan, UKM Medical Molecular Biology Institute, UKM Medical Centre, 8 Russia University Kebangsaan Malaysia, Kuala Lumpur, Malaysia, Division of Genetics and Molecular Biology, Faculty of Science, Antoine Danchin, Institute of Biological Sciences, University of Malaya, Kuala Lumpur, Malaysia Institut de Cardiométabolisme et Nutrition (ICAN), France Streptomyces colonosanans MUSC 93JT, a novel strain isolated from mangrove forest *Correspondence: Bey-Hing Goh soil located at Sarawak, Malaysia. The bacterium was noted to be Gram-positive and [email protected] to form light yellow aerial and vivid yellow substrate mycelium on ISP 2 agar.
    [Show full text]
  • The Hns Gene of Escherichia Coli Is Transcriptionally Down-Regulated by (P)Ppgpp
    microorganisms Article The hns Gene of Escherichia coli Is Transcriptionally Down-Regulated by (p)ppGpp Anna Brandi, Mara Giangrossi, Attilio Fabbretti and Maurizio Falconi * School of Biosciences and Veterinary Medicine, University of Camerino, 62032 Camerino, Italy; [email protected] (A.B.); [email protected] (M.G.); [email protected] (A.F.) * Correspondence: [email protected]; Tel.: +39-0737-403274 Received: 25 August 2020; Accepted: 8 October 2020; Published: 10 October 2020 Abstract: Second messenger nucleotides, such as guanosine penta- or tetra-phosphate, commonly referred to as (p)ppGpp, are powerful signaling molecules, used by all bacteria to fine-tune cellular metabolism in response to nutrient availability. Indeed, under nutritional starvation, accumulation of (p)ppGpp reduces cell growth, inhibits stable RNAs synthesis, and selectively up- or down- regulates the expression of a large number of genes. Here, we show that the E. coli hns promoter responds to intracellular level of (p)ppGpp. hns encodes the DNA binding protein H-NS, one of the major components of bacterial nucleoid. Currently, H-NS is viewed as a global regulator of transcription in an environment-dependent mode. Combining results from relA (ppGpp synthetase) and spoT (ppGpp synthetase/hydrolase) null mutants with those from an inducible plasmid encoded RelA system, we have found that hns expression is inversely correlated with the intracellular concentration of (p)ppGpp, particularly in exponential phase of growth. Furthermore, we have reproduced in an in vitro system the observed in vivo (p)ppGpp-mediated transcriptional repression of hns promoter. Electrophoretic mobility shift assays clearly demonstrated that this unusual nucleotide negatively affects the stability of RNA polymerase-hns promoter complex.
    [Show full text]
  • Anticancer Drug Discovery from Microbial Sources: the Unique Mangrove Streptomycetes
    molecules Review Anticancer Drug Discovery from Microbial Sources: The Unique Mangrove Streptomycetes Jodi Woan-Fei Law 1, Lydia Ngiik-Shiew Law 2, Vengadesh Letchumanan 1 , Loh Teng-Hern Tan 1, Sunny Hei Wong 3, Kok-Gan Chan 4,5,* , Nurul-Syakima Ab Mutalib 6,* and Learn-Han Lee 1,* 1 Novel Bacteria and Drug Discovery (NBDD) Research Group, Microbiome and Bioresource Research Strength, Jeffrey Cheah School of Medicine and Health Sciences, Monash University Malaysia, Bandar Sunway 47500, Selangor Darul Ehsan, Malaysia; [email protected] (J.W.-F.L.); [email protected] (V.L.); [email protected] (L.T.-H.T.) 2 Monash Credentialed Pharmacy Clinical Educator, Faculty of Pharmacy and Pharmaceutical Sciences, Monash University, 381 Royal Parade, Parkville 3052, VIC, Australia; [email protected] 3 Li Ka Shing Institute of Health Sciences, Department of Medicine and Therapeutics, The Chinese University of Hong Kong, Shatin, Hong Kong, China; [email protected] 4 Division of Genetics and Molecular Biology, Institute of Biological Sciences, Faculty of Science, University of Malaya, Kuala Lumpur 50603, Malaysia 5 International Genome Centre, Jiangsu University, Zhenjiang 212013, China 6 UKM Medical Molecular Biology Institute (UMBI), UKM Medical Centre, Universiti Kebangsaan Malaysia, Kuala Lumpur 56000, Malaysia * Correspondence: [email protected] (K.-G.C.); [email protected] (N.-S.A.M.); [email protected] (L.-H.L.) Academic Editor: Owen M. McDougal Received: 8 October 2020; Accepted: 13 November 2020; Published: 17 November 2020 Abstract: Worldwide cancer incidence and mortality have always been a concern to the community. The cancer mortality rate has generally declined over the years; however, there is still an increased mortality rate in poorer countries that receives considerable attention from healthcare professionals.
    [Show full text]
  • Improved Taxonomy of the Genus Streptomyces
    UNIVERSITEIT GENT Faculteit Wetenschappen Vakgroep Biochemie, Fysiologie & Microbiologie Laboratorium voor Microbiologie Improved taxonomy of the genus Streptomyces Benjamin LANOOT Scriptie voorgelegd tot het behalen van de graad van Doctor in de Wetenschappen (Biochemie) Promotor: Prof. Dr. ir. J. Swings Co-promotor: Dr. M. Vancanneyt Academiejaar 2004-2005 FACULTY OF SCIENCES ____________________________________________________________ DEPARTMENT OF BIOCHEMISTRY, PHYSIOLOGY AND MICROBIOLOGY UNIVERSITEIT LABORATORY OF MICROBIOLOGY GENT IMPROVED TAXONOMY OF THE GENUS STREPTOMYCES DISSERTATION Submitted in fulfilment of the requirements for the degree of Doctor (Ph D) in Sciences, Biochemistry December 2004 Benjamin LANOOT Promotor: Prof. Dr. ir. J. SWINGS Co-promotor: Dr. M. VANCANNEYT 1: Aerial mycelium of a Streptomyces sp. © Michel Cavatta, Academy de Lyon, France 1 2 2: Streptomyces coelicolor colonies © John Innes Centre 3: Blue haloes surrounding Streptomyces coelicolor colonies are secreted 3 4 actinorhodin (an antibiotic) © John Innes Centre 4: Antibiotic droplet secreted by Streptomyces coelicolor © John Innes Centre PhD thesis, Faculty of Sciences, Ghent University, Ghent, Belgium. Publicly defended in Ghent, December 9th, 2004. Examination Commission PROF. DR. J. VAN BEEUMEN (ACTING CHAIRMAN) Faculty of Sciences, University of Ghent PROF. DR. IR. J. SWINGS (PROMOTOR) Faculty of Sciences, University of Ghent DR. M. VANCANNEYT (CO-PROMOTOR) Faculty of Sciences, University of Ghent PROF. DR. M. GOODFELLOW Department of Agricultural & Environmental Science University of Newcastle, UK PROF. Z. LIU Institute of Microbiology Chinese Academy of Sciences, Beijing, P.R. China DR. D. LABEDA United States Department of Agriculture National Center for Agricultural Utilization Research Peoria, IL, USA PROF. DR. R.M. KROPPENSTEDT Deutsche Sammlung von Mikroorganismen & Zellkulturen (DSMZ) Braunschweig, Germany DR.
    [Show full text]
  • 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.
    [Show full text]
  • Evolution of the Streptomycin and Viomycin Biosynthetic Clusters and Resistance Genes
    University of Warwick institutional repository: http://go.warwick.ac.uk/wrap A Thesis Submitted for the Degree of PhD at the University of Warwick http://go.warwick.ac.uk/wrap/2773 This thesis is made available online and is protected by original copyright. Please scroll down to view the document itself. Please refer to the repository record for this item for information to help you to cite it. Our policy information is available from the repository home page. Evolution of the streptomycin and viomycin biosynthetic clusters and resistance genes Paris Laskaris, B.Sc. (Hons.) A thesis submitted to the University of Warwick for the degree of Doctor of Philosophy. Department of Biological Sciences, University of Warwick, Coventry, CV4 7AL September 2009 Contents List of Figures ........................................................................................................................ vi List of Tables ....................................................................................................................... xvi Abbreviations ........................................................................................................................ xx Acknowledgements .............................................................................................................. xxi Declaration .......................................................................................................................... xxii Abstract .............................................................................................................................
    [Show full text]
  • 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.
    [Show full text]
  • Description of Streptomyces Dangxiongensis Sp. Nov
    Cronfa - Swansea University Open Access Repository _____________________________________________________________ This is an author produced version of a paper published in: International Journal of Systematic and Evolutionary Microbiology Cronfa URL for this paper: http://cronfa.swan.ac.uk/Record/cronfa50961 _____________________________________________________________ Paper: Zhang, B., Tang, S., Yang, R., Chen, X., Zhang, D., Zhang, W., Li, S., Chen, T., Liu, G. et. al. (2019). Streptomyces dangxiongensis sp. nov., isolated from soil of Qinghai-Tibet Plateau. International Journal of Systematic and Evolutionary Microbiology http://dx.doi.org/10.1099/ijsem.0.003550 _____________________________________________________________ This item is brought to you by Swansea University. Any person downloading material is agreeing to abide by the terms of the repository licence. Copies of full text items may be used or reproduced in any format or medium, without prior permission for personal research or study, educational or non-commercial purposes only. The copyright for any work remains with the original author unless otherwise specified. The full-text must not be sold in any format or medium without the formal permission of the copyright holder. Permission for multiple reproductions should be obtained from the original author. Authors are personally responsible for adhering to copyright and publisher restrictions when uploading content to the repository. http://www.swansea.ac.uk/library/researchsupport/ris-support/ Streptomyces dangxiongensis sp. nov., isolated from soil of Qinghai-Tibet Plateau Binglin Zhang1,2,3, Shukun Tang4, Ruiqi Yang1,3, Ximing Chen1,3, Dongming Zhang1, Wei Zhang1,3, Shiweng Li5, Tuo Chen2,3, Guangxiu Liu1,3, Paul Dyson6 1 Key Laboratory of Desert and Desertification, Northwest Institute of Eco-Environment and Resources, Chinese Academy of Sciences, Lanzhou 730000, China.
    [Show full text]
  • 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,
    [Show full text]
  • 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.
    [Show full text]
  • Genomic and Phylogenomic Insights Into the Family Streptomycetaceae Lead to Proposal of Charcoactinosporaceae Fam. Nov. and 8 No
    bioRxiv preprint doi: https://doi.org/10.1101/2020.07.08.193797; this version posted July 8, 2020. 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. 1 Genomic and phylogenomic insights into the family Streptomycetaceae 2 lead to proposal of Charcoactinosporaceae fam. nov. and 8 novel genera 3 with emended descriptions of Streptomyces calvus 4 Munusamy Madhaiyan1, †, * Venkatakrishnan Sivaraj Saravanan2, † Wah-Seng See-Too3, † 5 1Temasek Life Sciences Laboratory, 1 Research Link, National University of Singapore, 6 Singapore 117604; 2Department of Microbiology, Indira Gandhi College of Arts and Science, 7 Kathirkamam 605009, Pondicherry, India; 3Division of Genetics and Molecular Biology, 8 Institute of Biological Sciences, Faculty of Science, University of Malaya, Kuala Lumpur, 9 Malaysia 10 *Corresponding author: Temasek Life Sciences Laboratory, 1 Research Link, National 11 University of Singapore, Singapore 117604; E-mail: [email protected] 12 †All these authors have contributed equally to this work 13 Abstract 14 Streptomycetaceae is one of the oldest families within phylum Actinobacteria and it is large and 15 diverse in terms of number of described taxa. The members of the family are known for their 16 ability to produce medically important secondary metabolites and antibiotics. In this study, 17 strains showing low 16S rRNA gene similarity (<97.3 %) with other members of 18 Streptomycetaceae were identified and subjected to phylogenomic analysis using 33 orthologous 19 gene clusters (OGC) for accurate taxonomic reassignment resulted in identification of eight 20 distinct and deeply branching clades, further average amino acid identity (AAI) analysis showed 1 bioRxiv preprint doi: https://doi.org/10.1101/2020.07.08.193797; this version posted July 8, 2020.
    [Show full text]
  • 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.
    [Show full text]