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Chitin synthases from Saprolegnia are involved in tip growth and represent a potential target for anti-oomycete drugs.
PLoS Pathog. 2010 Aug 26; 6(8):e1001070.PP

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.

Authors+Show Affiliations

Division of Glycoscience, School of Biotechnology, Royal Institute of Technology, AlbaNova University Centre, Stockholm, Sweden.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

20865175

Citation

Guerriero, Gea, et al. "Chitin Synthases From Saprolegnia Are Involved in Tip Growth and Represent a Potential Target for Anti-oomycete Drugs." PLoS Pathogens, vol. 6, no. 8, 2010, pp. e1001070.
Guerriero G, Avino M, Zhou Q, et al. Chitin synthases from Saprolegnia are involved in tip growth and represent a potential target for anti-oomycete drugs. PLoS Pathog. 2010;6(8):e1001070.
Guerriero, G., Avino, M., Zhou, Q., Fugelstad, J., Clergeot, P. H., & Bulone, V. (2010). Chitin synthases from Saprolegnia are involved in tip growth and represent a potential target for anti-oomycete drugs. PLoS Pathogens, 6(8), e1001070. https://doi.org/10.1371/journal.ppat.1001070
Guerriero G, et al. Chitin Synthases From Saprolegnia Are Involved in Tip Growth and Represent a Potential Target for Anti-oomycete Drugs. PLoS Pathog. 2010 Aug 26;6(8):e1001070. PubMed PMID: 20865175.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Chitin synthases from Saprolegnia are involved in tip growth and represent a potential target for anti-oomycete drugs. AU - Guerriero,Gea, AU - Avino,Mariano, AU - Zhou,Qi, AU - Fugelstad,Johanna, AU - Clergeot,Pierre-Henri, AU - Bulone,Vincent, Y1 - 2010/08/26/ PY - 2010/03/08/received PY - 2010/07/26/accepted PY - 2010/9/25/entrez PY - 2010/9/25/pubmed PY - 2010/12/14/medline SP - e1001070 EP - e1001070 JF - PLoS pathogens JO - PLoS Pathog. VL - 6 IS - 8 N2 - 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. SN - 1553-7374 UR - https://www.unboundmedicine.com/medline/citation/20865175/Chitin_synthases_from_Saprolegnia_are_involved_in_tip_growth_and_represent_a_potential_target_for_anti_oomycete_drugs_ L2 - http://dx.plos.org/10.1371/journal.ppat.1001070 DB - PRIME DP - Unbound Medicine ER -