Please use this identifier to cite or link to this item: https://hdl.handle.net/2440/89262
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Type: Journal article
Title: Chitin synthases from saprolegnia are involved in tip growth and represent a potential target for anti-Oomycete drugs
Author: Guerriero, G.
Avino, M.
Zhou, Q.
Fugelstad, J.
Clergeot, P.H.
Bulone, V.
Citation: PLoS Pathogens, 2010; 6(8):e1001070-1-e1001070-12
Publisher: Public Library of Science
Issue Date: 2010
ISSN: 1553-7374
1553-7366
Editor: Kamoun, S.
Statement of
Responsibility: 
Gea Guerriero, Mariano Avino, Qi Zhou, Johanna Fugelstad, Pierre-Henri Clergeot, Vincent Bulone
Abstract: Oomycetes represent some of the most devastating plant and animal pathogens. Typical examples are Phytophthora infestans, which causes potato and tomato late blight, and Saprolegnia parasitica, responsible for fish diseases. Despite the economical and environmental importance of oomycete diseases, their control is difficult, particularly in the aquaculture industry. Carbohydrate synthases are vital for hyphal growth and represent interesting targets for tackling the pathogens. The existence of 2 different chitin synthase genes (SmChs1 and SmChs2) in Saprolegnia monoica was demonstrated using bioinformatics and molecular biology approaches. The function of SmCHS2 was unequivocally demonstrated by showing its catalytic activity in vitro after expression in Pichia pastoris. The recombinant SmCHS1 protein did not exhibit any activity in vitro, suggesting that it requires other partners or effectors to be active, or that it is involved in a different process than chitin biosynthesis. Both proteins contained N-terminal Microtubule Interacting and Trafficking domains, which have never been reported in any other known carbohydrate synthases. These domains are involved in protein recycling by endocytosis. Enzyme kinetics revealed that Saprolegnia chitin synthases are competitively inhibited by nikkomycin Z and quantitative PCR showed that their expression is higher in presence of the inhibitor. The use of nikkomycin Z combined with microscopy showed that chitin synthases are active essentially at the hyphal tips, which burst in the presence of the inhibitor, leading to cell death. S. parasitica was more sensitive to nikkomycin Z than S. monoica. In conclusion, chitin synthases with species-specific characteristics are involved in tip growth in Saprolegnia species and chitin is vital for the micro-organisms despite its very low abundance in the cell walls. Chitin is most likely synthesized transiently at the apex of the cells before cellulose, the major cell wall component in oomycetes. Our results provide important fundamental information on cell wall biogenesis in economically important species, and demonstrate the potential of targeting oomycete chitin synthases for disease control.
Keywords: Saprolegnia
Chitin Synthase
Aminoglycosides
Microscopy, Electron
Antifungal Agents
Blotting, Southern
Reverse Transcriptase Polymerase Chain Reaction
Phylogeny
Amino Acid Sequence
Molecular Sequence Data
Rights: © 2010 Guerriero et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
DOI: 10.1371/journal.ppat.1001070
Published version: http://dx.doi.org/10.1371/journal.ppat.1001070
Appears in Collections:Agriculture, Food and Wine publications
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