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Exploiting yeast cell wall for high throughput screening of antimicrobial agents

   
Project

QLK3-2000-01537

Cell factory area

3.3.1

EU Contribution

1 232 275 Euro

Duration

36 months

Type

Research project

Starting date

01-11-2000

Keywords
high throughput screening
Saccharomyces cerevisiae
yeast cell wall
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ABSTRACT
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The main objective of this proposal is to exploit the molecular knowledge of the Saccharomyces cerevisiae cell wall for high throughput screening of antimicrobial agents. To this end, a consortium of 10 laboratories (one company, one SME and 8 public research institutes) will convert the molecular data on essential gene targets involved in cell wall cross-linking, remodelling and chitin pathways into assays amenable for drug-discovery programmes and to apply genomics, proteomics and bio-informatics, to identify new targets through the characterisation of the cell wall compensatory mechanism which is induced when the cell wall is weakened by drug treatment, stress, or mutations. The deliverables will be the production of number of assays for high throughput screenings, and the goal is to find novel antifungal molecules acting on the designed targets.

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OBJECTIVES
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The scientific objectives of this proposal are:

  • To characterise the cross-linking and remodelling pathways which are the basis for the modular structure of the cell wall in pathogen and non-pathogen fungi, using the yeast S. cerevisiae as the model system.
  • To identify by an integrated approach involving genomics, proteomics and bio-informatics, the determinants of the cell wall compensatory mechanism which is induced when the cell wall is weakened by various treatments.
  • The technological objective is to provide assays from selected cell wall targets for high throughput screening.
  • The socio-economic impact of the project is to speed up the search of novel effective antifungal drugs. These objectives will be achieved by a strong commitment between molecular and cellular identification of cell wall targets and their biochemical characterisation to design assays allowing development of drug-discovery programmes.

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DESCRIPTION OF THE WORK
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The objectives will be reached by an integrated programme which will be divided into four workpackages.

Workpackage I will deal with the molecular and biochemical characterisation of cell wall cross-linking and remodelling pathways, identifying the different transglycosidases, beta-1,3 and 1,6 glucantransferase and chitinolytic/transglycosidic enzymes which catalyse critical steps of linkage between the 3 essential macromolecules which compose the cell wall. This characterisation will be followed by the design of assays for high throughput screening.

Workpackage II will deal with a detailed molecular and biochemical characterisation of the complex chitin synthesis pathway. This work will result in the setting-up of a novel chitin synthase assay which is highly effective for high throughput screening since it will take into account all newly regulatory proteins implicated in control of chitin synthesis.

Workpackage III will be devoted to the identification of new potential cell wall targets through a global genomic and proteomic analysis of the cell wall compensatory mechanism which is activated when the cell wall is weakened by drugs treatment, stress or mutations.

Workpackage IV will deal with the administrative, financial and report-writing aspects. All interesting, non-confidential data arising during the work will be communicated through a WEB site.

To raise to this challenge, eight leading academic research groups in the fields of the biochemistry and molecular biology of the fungal cell wall, genomics, proteomics, and bio-informatics will pool their expertise and skills, to increase our knowledge on the molecular and biochemical mechanisms involved in cell wall assembly, and to provide industry with reliable assays on well-characterised cell targets for high throughput screening of antimicrobial agents. The discovery of an antifungal sample from at least two of the selected targets is expected at the end of the project.

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DELIVERABLES
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  • Provide specific assays of well-dedicated cell wall enzymes for high throughput screening (HTS).
  • Evaluate enzymes of the cross-linking and remodelling pathways as novel putative targets, and identify new targets from genomic and proteomic analysis (Month 36).
  • Identify novel antifungal activities.
  • Provide progress reports every 6 months

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CONSORTIUM
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COORDINATOR
  Jean-Marie François
Department Genie Biochimique et Alimentaire
Institut National des Sciences Appliquées
31077 Toulouse, France
Tel: +33 561559492
Fax: +33 56159492
fran_jm@insa-tlse.fr

PARTNERS
  Laura Popolo
Universita Degli Studi di Milano
Dipt Fisiologia e Biochemica generali
Via Celoria 26
20133 Milan, Italy
Tel: +39 270644808
Fax: +39 270632811
laura.popolo@unimi.it

Cesar Roncero
Departamento de Microbiologia
Eidificio Departamental H-219
Avda Camp Charros S/N
37007 Salamanca, Spain
Tel: +34 923294733
crm@gugu.usal.es

Lucia Carrano
Microbial Technologies
Biosearch italia Spa
21040 Gerenzano, Italy
Tel: +39 0296474432
Fax: +39 0296474238
lcarrano@biosearch.it

Jean-Luc Zundel
Aventis Crop Science SA
Centre de Recherche de la Dargoire
14-20 rue Pierre Baizet
69009 Lyon, France
Tel: +33 472852823
Fax: +33 472852297
jean-luc.zundel@aventis.com

Graham Gooday
Deparment of Molec. Cell Biol
University of Aberdeen
AB25 2ZD Aberdeen, United Kingdom
Tel: +44 1224273147
Fax: +44 1224273144
g.w.gooday@abdn.ac.uk

Widmar Tanner
Lehstuhl für Zell Biologie und
Planzenphysiologie
Universität Regensburg
93040 Regensburg, Germany
Tel: +49 9419433018
Fax: +49 9419433352
widmar.tanner@biologie.uni-regensburg.de

Francisco Del Rey
Dept de Microbiologica y Genetica
Universidad de Salamanca
Campus Miguel de Unamuno
37007 Salamanca, Spain
Tel: +34 923294675
Fax: +34 923224876
fdelrey@gugu.usal.es

Joerg Hoheisel
Functional Genome Analysis
Deutsches Krebsforschungscentrum
Im Neuenheimer Feld 506
69120 Heidelberg, Germany
Tel: +49 6221424680
Fax: +49 6221424682
j.hoheisel@dkfz-heidelberg.de
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