title: A Single Protein S-acyl Transferase Acts through Diverse Substrates to Determine Cryptococcal Morphology, Stress Tolerance, and Pathogenic Outcome.
abstract: Cryptococcus neoformans is an opportunistic yeast that kills over 625,000 people yearly through lethal meningitis. Host phagocytes serve as the first line of defense against this pathogen, but fungal engulfment and subsequent intracellular proliferation also correlate with poor patient outcome. Defining the interactions of this facultative intracellular pathogen with host phagocytes is key to understanding the latter's opposing roles in infection and how they contribute to fungal latency, dissemination, and virulence. We used high-content imaging and a human monocytic cell line to screen 1,201 fungal mutants for strains with altered host interactions and identified multiple genes that influence fungal adherence and phagocytosis. One of these genes was PFA4, which encodes a protein S-acyl transferase (PAT), one of a family of DHHC domain-containing proteins that catalyzes lipid modification of proteins. Deletion of PFA4 caused dramatic defects in cryptococcal morphology, stress tolerance, and virulence. Bioorthogonal palmitoylome-profiling identified Pfa4-specific protein substrates involved in cell wall synthesis, signal transduction, and membrane trafficking responsible for these phenotypic alterations. We demonstrate that a single PAT is responsible for the modification of a subset of proteins that are critical in cryptococcal pathogenesis. Since several of these palmitoylated substrates are conserved in other pathogenic fungi, protein palmitoylation represents a potential avenue for new antifungal therapeutics.
authors.affiliation: Department of Molecular Microbiology, Washington University School of Medicine, St. Louis, Missouri, United States of America.
authors.forename: Felipe H
authors.initials: FH
authors.lastname: Santiago-Tirado
authors.affiliation: Laboratory of Chemical Biology and Microbial Pathogenesis, The Rockefeller University, New York, New York, United States of America.
authors.forename: Tao
authors.initials: T
authors.lastname: Peng
authors.affiliation: Department of Molecular Microbiology, Washington University School of Medicine, St. Louis, Missouri, United States of America.
authors.forename: Meng
authors.initials: M
authors.lastname: Yang
authors.affiliation: Laboratory of Chemical Biology and Microbial Pathogenesis, The Rockefeller University, New York, New York, United States of America.
authors.forename: Howard C
authors.initials: HC
authors.lastname: Hang
authors.affiliation: Department of Molecular Microbiology, Washington University School of Medicine, St. Louis, Missouri, United States of America.
authors.forename: Tamara L
authors.initials: TL
authors.lastname: Doering
mesh_terms: D000215:Acylation; D000217:Acyltransferases / Q000235:genetics / Q000378:metabolism*; D002448:Cell Adhesion; D002460:Cell Line; D002473:Cell Wall / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D003453:Cryptococcosis / Q000276:immunology / Q000378:metabolism* / Q000382:microbiology / Q000473:pathology; D003455:Cryptococcus neoformans / Q000166:cytology / Q000201:enzymology / Q000472:pathogenicity / Q000502:physiology*; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D017353:Gene Deletion; D054884:Host-Pathogen Interactions; D006801:Humans; D016919:Meningitis, Cryptococcal / Q000276:immunology / Q000378:metabolism / Q000382:microbiology / Q000473:pathology; D050296:Microbial Viability; D009000:Monocytes / Q000276:immunology / Q000378:metabolism / Q000382:microbiology* / Q000473:pathology; D009154:Mutation; D010587:Phagocytosis; D011499:Protein Processing, Post-Translational; D015398:Signal Transduction; D013312:Stress, Physiological; D013379:Substrate Specificity; D014774:Virulence; D017735:Virus Latency
chemical_list: D005656:Fungal Proteins; D000217:Acyltransferases