title: RNAi-Based Functional Genomics Identifies New Virulence Determinants in Mucormycosis.
abstract: Mucorales are an emerging group of human pathogens that are responsible for the lethal disease mucormycosis. Unfortunately, functional studies on the genetic factors behind the virulence of these organisms are hampered by their limited genetic tractability, since they are reluctant to classical genetic tools like transposable elements or gene mapping. Here, we describe an RNAi-based functional genomic platform that allows the identification of new virulence factors through a forward genetic approach firstly described in Mucorales. This platform contains a whole-genome collection of Mucor circinelloides silenced transformants that presented a broad assortment of phenotypes related to the main physiological processes in fungi, including virulence, hyphae morphology, mycelial and yeast growth, carotenogenesis and asexual sporulation. Selection of transformants with reduced virulence allowed the identification of mcplD, which encodes a Phospholipase D, and mcmyo5, encoding a probably essential cargo transporter of the Myosin V family, as required for a fully virulent phenotype of M. circinelloides. Knock-out mutants for those genes showed reduced virulence in both Galleria mellonella and Mus musculus models, probably due to a delayed germination and polarized growth within macrophages. This study provides a robust approach to study virulence in Mucorales and as a proof of concept identified new virulence determinants in M. circinelloides that could represent promising targets for future antifungal therapies.
authors.affiliation: Departmento de Genética y Microbiología, Facultad de Biología, Universidad de Murcia, Spain.
authors.forename: Trung Anh
authors.identifier: 0000-0002-4220-9078
authors.initials: TA
authors.lastname: Trieu
authors.affiliation: Departmento de Genética y Microbiología, Facultad de Biología, Universidad de Murcia, Spain.
authors.forename: María Isabel
authors.identifier: 0000-0002-3560-6155
authors.initials: MI
authors.lastname: Navarro-Mendoza
authors.affiliation: Departmento de Genética y Microbiología, Facultad de Biología, Universidad de Murcia, Spain.
authors.forename: Carlos
authors.identifier: 0000-0003-1384-3608
authors.initials: C
authors.lastname: Pérez-Arques
authors.affiliation: Unidad de Microbiología, Universitat Rovira i Virgili, IISPV, Tarragona, Spain.
authors.forename: Marta
authors.initials: M
authors.lastname: Sanchis
authors.affiliation: Unidad de Microbiología, Universitat Rovira i Virgili, IISPV, Tarragona, Spain.
authors.forename: Javier
authors.identifier: 0000-0002-0765-6403
authors.initials: J
authors.lastname: Capilla
authors.affiliation: Unidad de Microbiología, Universitat Rovira i Virgili, IISPV, Tarragona, Spain.
authors.forename: Patricia
authors.identifier: 0000-0003-4192-6291
authors.initials: P
authors.lastname: Navarro-Rodriguez
authors.affiliation: Unidad de Microbiología, Universitat Rovira i Virgili, IISPV, Tarragona, Spain.
authors.forename: Loida
authors.initials: L
authors.lastname: Lopez-Fernandez
authors.affiliation: Departmento de Genética y Microbiología, Facultad de Biología, Universidad de Murcia, Spain.
authors.forename: Santiago
authors.initials: S
authors.lastname: Torres-Martínez
authors.affiliation: Departmento de Genética y Microbiología, Facultad de Biología, Universidad de Murcia, Spain.
authors.forename: Victoriano
authors.initials: V
authors.lastname: Garre
authors.affiliation: Departmento de Genética y Microbiología, Facultad de Biología, Universidad de Murcia, Spain.
authors.forename: Rosa María
authors.initials: RM
authors.lastname: Ruiz-Vázquez
authors.affiliation: Departmento de Genética y Microbiología, Facultad de Biología, Universidad de Murcia, Spain.
authors.forename: Francisco E
authors.identifier: 0000-0003-3762-5971
authors.initials: FE
authors.lastname: Nicolás
mesh_terms: D000818:Animals; D000935:Antifungal Agents / Q000494:pharmacology; D026141:Drug Resistance, Multiple, Fungal; D005656:Fungal Proteins / Q000235:genetics*; D007814:Larva / Q000382:microbiology*; D008264:Macrophages / Q000382:microbiology; D008297:Male; D051379:Mice; D009036:Moths / Q000382:microbiology*; D009089:Mucor / Q000235:genetics / Q000472:pathogenicity*; D009091:Mucormycosis / Q000473:pathology* / Q000821:virology; D024701:Myosin Type V / Q000235:genetics*; D010739:Phospholipase D / Q000235:genetics*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D037521:Virulence Factors / Q000235:genetics*
chemical_list: D000935:Antifungal Agents; D005656:Fungal Proteins; D034741:RNA, Small Interfering; D037521:Virulence Factors; D010739:Phospholipase D; D024701:Myosin Type V