title: Impact of the lectin chaperone calnexin on the stress response, virulence and proteolytic secretome of the fungal pathogen Aspergillus fumigatus.
abstract: Calnexin is a membrane-bound lectin chaperone in the endoplasmic reticulum (ER) that is part of a quality control system that promotes the accurate folding of glycoproteins entering the secretory pathway. We have previously shown that ER homeostasis is important for virulence of the human fungal pathogen Aspergillus fumigatus, but the contribution of calnexin has not been explored. Here, we determined the extent to which A. fumigatus relies on calnexin for growth under conditions of environmental stress and for virulence. The calnexin gene, clxA, was deleted from A. fumigatus and complemented by reconstitution with the wild type gene. Loss of clxA altered the proteolytic secretome of the fungus, but had no impact on growth rates in either minimal or complex media at 37°C. However, the ΔclxA mutant was growth impaired at temperatures above 42°C and was hypersensitive to acute ER stress caused by the reducing agent dithiothreitol. In contrast to wild type A. fumigatus, ΔclxA hyphae were unable to grow when transferred to starvation medium. In addition, depleting the medium of cations by chelation prevented ΔclxA from sustaining polarized hyphal growth, resulting in blunted hyphae with irregular morphology. Despite these abnormal stress responses, the ΔclxA mutant remained virulent in two immunologically distinct models of invasive aspergillosis. These findings demonstrate that calnexin functions are needed for growth under conditions of thermal, ER and nutrient stress, but are dispensable for surviving the stresses encountered in the host environment.
authors.affiliation: Department of Pathology & Laboratory Medicine, University of Cincinnati College of Medicine, Cincinnati, Ohio, United States of America.
authors.forename: Margaret V
authors.initials: MV
authors.lastname: Powers-Fletcher
authors.forename: Kalyani
authors.initials: K
authors.lastname: Jambunathan
authors.forename: Jordan L
authors.initials: JL
authors.lastname: Brewer
authors.forename: Karthik
authors.initials: K
authors.lastname: Krishnan
authors.forename: Xizhi
authors.initials: X
authors.lastname: Feng
authors.forename: Amit K
authors.initials: AK
authors.lastname: Galande
authors.forename: David S
authors.initials: DS
authors.lastname: Askew
mesh_terms: D000818:Animals; D001232:Aspergillus fumigatus / Q000378:metabolism*; D037281:Calnexin / Q000737:chemistry* / Q000235:genetics / Q000502:physiology*; D002412:Cations; D003470:Culture Media / Q000494:pharmacology; D017931:DNA Primers / Q000235:genetics; D004721:Endoplasmic Reticulum / Q000378:metabolism; D059865:Endoplasmic Reticulum Stress; D005260:Female; D005656:Fungal Proteins / Q000235:genetics / Q000502:physiology*; D015966:Gene Expression Regulation, Fungal; D007166:Immunosuppressive Agents / Q000627:therapeutic use; D037102:Lectins / Q000737:chemistry*; D051379:Mice; D016133:Polymerase Chain Reaction / Q000379:methods; D017510:Protein Folding; D013696:Temperature; D014774:Virulence
chemical_list: D002412:Cations; D003470:Culture Media; D017931:DNA Primers; D005656:Fungal Proteins; D007166:Immunosuppressive Agents; D037102:Lectins; D037281:Calnexin