title: Neurospora crassa transcriptomics reveals oxidative stress and plasma membrane homeostasis biology genes as key targets in response to chitosan.
abstract: Chitosan is a natural polymer with antimicrobial activity. Chitosan causes plasma membrane permeabilization and induction of intracellular reactive oxygen species (ROS) in Neurospora crassa. We have determined the transcriptional profile of N. crassa to chitosan and identified the main gene targets involved in the cellular response to this compound. Global network analyses showed membrane, transport and oxidoreductase activity as key nodes affected by chitosan. Activation of oxidative metabolism indicates the importance of ROS and cell energy together with plasma membrane homeostasis in N. crassa response to chitosan. Deletion strain analysis of chitosan susceptibility pointed NCU03639 encoding a class 3 lipase, involved in plasma membrane repair by lipid replacement, and NCU04537 a MFS monosaccharide transporter related to assimilation of simple sugars, as main gene targets of chitosan. NCU10521, a glutathione S-transferase-4 involved in the generation of reducing power for scavenging intracellular ROS is also a determinant chitosan gene target. Ca(2+) increased tolerance to chitosan in N. crassa. Growth of NCU10610 (fig 1 domain) and SYT1 (a synaptotagmin) deletion strains was significantly increased by Ca(2+) in the presence of chitosan. Both genes play a determinant role in N. crassa membrane homeostasis. Our results are of paramount importance for developing chitosan as an antifungal.
authors.affiliation: Laboratory of Plant Pathology, Multidisciplinary Institute for Environmental Studies (MIES) Ramon Margalef, Department of Marine Sciences and Applied Biology, University of Alicante, E-03080 Alicante, Spain. federico.lopez@ua.es lv.lopez@ua.es.
authors.forename: Federico
authors.initials: F
authors.lastname: Lopez-Moya
authors.affiliation: Department of Plant and Microbial Biology, University of California, Berkeley CA, 94720-3120 USA. djkowbel@berkeley.edu lglass@berkeley.edu.
authors.forename: David
authors.initials: D
authors.lastname: Kowbel
authors.affiliation: Statistics and Operation Research Department, University of Alicante, E-03080 Alicante, Spain. mj.nueda@ua.es.
authors.forename: Maria José
authors.initials: MJ
authors.lastname: Nueda
authors.affiliation: Department of Plant and Microbial Biology, University of California, Berkeley CA, 94720-3120 USA. djkowbel@berkeley.edu lglass@berkeley.edu.
authors.forename: Javier
authors.initials: J
authors.lastname: Palma-Guerrero
authors.affiliation: Department of Plant and Microbial Biology, University of California, Berkeley CA, 94720-3120 USA. djkowbel@berkeley.edu lglass@berkeley.edu.
authors.forename: N Louise
authors.initials: NL
authors.lastname: Glass
authors.affiliation: Laboratory of Plant Pathology, Multidisciplinary Institute for Environmental Studies (MIES) Ramon Margalef, Department of Marine Sciences and Applied Biology, University of Alicante, E-03080 Alicante, Spain. federico.lopez@ua.es lv.lopez@ua.es.
authors.forename: Luis Vicente
authors.initials: LV
authors.lastname: Lopez-Llorca
mesh_terms: D000935:Antifungal Agents / Q000494:pharmacology*; D002118:Calcium / Q000502:physiology; D002462:Cell Membrane / Q000378:metabolism*; D048271:Chitosan / Q000494:pharmacology*; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D020869:Gene Expression Profiling; D015966:Gene Expression Regulation, Fungal / Q000187:drug effects; D063990:Gene Ontology; D005800:Genes, Fungal; D006706:Homeostasis; D008826:Microbial Sensitivity Tests; D058977:Molecular Sequence Annotation; D009492:Neurospora crassa / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D017382:Reactive Oxygen Species / Q000378:metabolism; D013172:Spores, Fungal / Q000378:metabolism; D059467:Transcriptome / Q000187:drug effects*
chemical_list: D000935:Antifungal Agents; D005656:Fungal Proteins; D017382:Reactive Oxygen Species; D048271:Chitosan; D002118:Calcium