A Nonredundant Phosphopantetheinyl Transferase, PptA, Is a Novel Antifungal Target That Directs Secondary Metabolite, Siderophore, and Lysine Biosynthesis in <italic toggle="yes">Aspergillus fumigatus</italic> and Is Critical for Pathogenicity

ABSTRACT Secondary metabolites are key mediators of virulence for many pathogens. Aspergillus fumigatus produces a vast array of these bioactive molecules, the biosynthesis of which is catalyzed by nonribosomal peptide synthetases (NRPSs) or polyketide synthases (PKSs). Both NRPSs and PKSs harbor ca...

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Autores principales:	Anna Johns, Daniel H. Scharf, Fabio Gsaller, Hella Schmidt, Thorsten Heinekamp, Maria Straßburger, Jason D. Oliver, Mike Birch, Nicola Beckmann, Katharine S. Dobb, Jane Gilsenan, Bharatkumar Rash, Elaine Bignell, Axel A. Brakhage, Michael J. Bromley
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2017
Materias:	Aspergillus fumigatus antifungal agents drug targets gliotoxin secondary metabolism virulence determinants Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/cccea62e55644309b3c3b177d1c49935
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spelling	oai:doaj.org-article:cccea62e55644309b3c3b177d1c499352021-11-15T15:51:44ZA Nonredundant Phosphopantetheinyl Transferase, PptA, Is a Novel Antifungal Target That Directs Secondary Metabolite, Siderophore, and Lysine Biosynthesis in <italic toggle="yes">Aspergillus fumigatus</italic> and Is Critical for Pathogenicity10.1128/mBio.01504-162150-7511https://doaj.org/article/cccea62e55644309b3c3b177d1c499352017-09-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01504-16https://doaj.org/toc/2150-7511ABSTRACT Secondary metabolites are key mediators of virulence for many pathogens. Aspergillus fumigatus produces a vast array of these bioactive molecules, the biosynthesis of which is catalyzed by nonribosomal peptide synthetases (NRPSs) or polyketide synthases (PKSs). Both NRPSs and PKSs harbor carrier domains that are primed for acceptance of secondary metabolic building blocks by a phosphopantetheinyl transferase (P-pant). The A. fumigatus P-pant PptA has been shown to prime the putative NRPS Pes1 in vitro and has an independent role in lysine biosynthesis; however, its role in global secondary metabolism and its impact on virulence has not been described. Here, we demonstrate that PptA has a nonredundant role in the generation of the vast majority of detectable secondary metabolites in A. fumigatus, including the immunomodulator gliotoxin, the siderophores triacetylfusarinine C (TAFC) and ferricrocin (FC), and dihydroxy naphthalene (DHN)-melanin. We show that both the lysine and iron requirements of a pptA null strain exceed those freely available in mammalian tissues and that loss of PptA renders A. fumigatus avirulent in both insect and murine infection models. Since PptA lacks similarity to its mammalian orthologue, we assert that the combined role of this enzyme in both primary and secondary metabolism, encompassing multiple virulence determinants makes it a very promising antifungal drug target candidate. We further exemplify this point with a high-throughput fluorescence polarization assay that we developed to identify chemical inhibitors of PptA function that have antifungal activity. IMPORTANCE Fungal diseases are estimated to kill between 1.5 and 2 million people each year, which exceeds the global mortality estimates for either tuberculosis or malaria. Only four classes of antifungal agents are available to treat invasive fungal infections, and all suffer pharmacological shortcomings, including toxicity, drug-drug interactions, and poor bioavailability. There is an urgent need to develop a new class of drugs that operate via a novel mechanism of action. We have identified a potential drug target, PptA, in the fungal pathogen Aspergillus fumigatus. PptA is required to synthesize the immunotoxic compound gliotoxin, DHN-melanin, which A. fumigatus employs to evade detection by host cells, the amino acid lysine, and the siderophores TAFC and FC, which A. fumigatus uses to scavenge iron. We show that strains lacking the PptA enzyme are unable to establish an infection, and we present a method which we use to identify novel antifungal drugs that inactivate PptA.Anna JohnsDaniel H. ScharfFabio GsallerHella SchmidtThorsten HeinekampMaria StraßburgerJason D. OliverMike BirchNicola BeckmannKatharine S. DobbJane GilsenanBharatkumar RashElaine BignellAxel A. BrakhageMichael J. BromleyAmerican Society for MicrobiologyarticleAspergillus fumigatusantifungal agentsdrug targetsgliotoxinsecondary metabolismvirulence determinantsMicrobiologyQR1-502ENmBio, Vol 8, Iss 4 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Aspergillus fumigatus antifungal agents drug targets gliotoxin secondary metabolism virulence determinants Microbiology QR1-502
spellingShingle	Aspergillus fumigatus antifungal agents drug targets gliotoxin secondary metabolism virulence determinants Microbiology QR1-502 Anna Johns Daniel H. Scharf Fabio Gsaller Hella Schmidt Thorsten Heinekamp Maria Straßburger Jason D. Oliver Mike Birch Nicola Beckmann Katharine S. Dobb Jane Gilsenan Bharatkumar Rash Elaine Bignell Axel A. Brakhage Michael J. Bromley A Nonredundant Phosphopantetheinyl Transferase, PptA, Is a Novel Antifungal Target That Directs Secondary Metabolite, Siderophore, and Lysine Biosynthesis in <italic toggle="yes">Aspergillus fumigatus</italic> and Is Critical for Pathogenicity
description	ABSTRACT Secondary metabolites are key mediators of virulence for many pathogens. Aspergillus fumigatus produces a vast array of these bioactive molecules, the biosynthesis of which is catalyzed by nonribosomal peptide synthetases (NRPSs) or polyketide synthases (PKSs). Both NRPSs and PKSs harbor carrier domains that are primed for acceptance of secondary metabolic building blocks by a phosphopantetheinyl transferase (P-pant). The A. fumigatus P-pant PptA has been shown to prime the putative NRPS Pes1 in vitro and has an independent role in lysine biosynthesis; however, its role in global secondary metabolism and its impact on virulence has not been described. Here, we demonstrate that PptA has a nonredundant role in the generation of the vast majority of detectable secondary metabolites in A. fumigatus, including the immunomodulator gliotoxin, the siderophores triacetylfusarinine C (TAFC) and ferricrocin (FC), and dihydroxy naphthalene (DHN)-melanin. We show that both the lysine and iron requirements of a pptA null strain exceed those freely available in mammalian tissues and that loss of PptA renders A. fumigatus avirulent in both insect and murine infection models. Since PptA lacks similarity to its mammalian orthologue, we assert that the combined role of this enzyme in both primary and secondary metabolism, encompassing multiple virulence determinants makes it a very promising antifungal drug target candidate. We further exemplify this point with a high-throughput fluorescence polarization assay that we developed to identify chemical inhibitors of PptA function that have antifungal activity. IMPORTANCE Fungal diseases are estimated to kill between 1.5 and 2 million people each year, which exceeds the global mortality estimates for either tuberculosis or malaria. Only four classes of antifungal agents are available to treat invasive fungal infections, and all suffer pharmacological shortcomings, including toxicity, drug-drug interactions, and poor bioavailability. There is an urgent need to develop a new class of drugs that operate via a novel mechanism of action. We have identified a potential drug target, PptA, in the fungal pathogen Aspergillus fumigatus. PptA is required to synthesize the immunotoxic compound gliotoxin, DHN-melanin, which A. fumigatus employs to evade detection by host cells, the amino acid lysine, and the siderophores TAFC and FC, which A. fumigatus uses to scavenge iron. We show that strains lacking the PptA enzyme are unable to establish an infection, and we present a method which we use to identify novel antifungal drugs that inactivate PptA.
format	article
author	Anna Johns Daniel H. Scharf Fabio Gsaller Hella Schmidt Thorsten Heinekamp Maria Straßburger Jason D. Oliver Mike Birch Nicola Beckmann Katharine S. Dobb Jane Gilsenan Bharatkumar Rash Elaine Bignell Axel A. Brakhage Michael J. Bromley
author_facet	Anna Johns Daniel H. Scharf Fabio Gsaller Hella Schmidt Thorsten Heinekamp Maria Straßburger Jason D. Oliver Mike Birch Nicola Beckmann Katharine S. Dobb Jane Gilsenan Bharatkumar Rash Elaine Bignell Axel A. Brakhage Michael J. Bromley
author_sort	Anna Johns
title	A Nonredundant Phosphopantetheinyl Transferase, PptA, Is a Novel Antifungal Target That Directs Secondary Metabolite, Siderophore, and Lysine Biosynthesis in <italic toggle="yes">Aspergillus fumigatus</italic> and Is Critical for Pathogenicity
title_short	A Nonredundant Phosphopantetheinyl Transferase, PptA, Is a Novel Antifungal Target That Directs Secondary Metabolite, Siderophore, and Lysine Biosynthesis in <italic toggle="yes">Aspergillus fumigatus</italic> and Is Critical for Pathogenicity
title_full	A Nonredundant Phosphopantetheinyl Transferase, PptA, Is a Novel Antifungal Target That Directs Secondary Metabolite, Siderophore, and Lysine Biosynthesis in <italic toggle="yes">Aspergillus fumigatus</italic> and Is Critical for Pathogenicity
title_fullStr	A Nonredundant Phosphopantetheinyl Transferase, PptA, Is a Novel Antifungal Target That Directs Secondary Metabolite, Siderophore, and Lysine Biosynthesis in <italic toggle="yes">Aspergillus fumigatus</italic> and Is Critical for Pathogenicity
title_full_unstemmed	A Nonredundant Phosphopantetheinyl Transferase, PptA, Is a Novel Antifungal Target That Directs Secondary Metabolite, Siderophore, and Lysine Biosynthesis in <italic toggle="yes">Aspergillus fumigatus</italic> and Is Critical for Pathogenicity
title_sort	nonredundant phosphopantetheinyl transferase, ppta, is a novel antifungal target that directs secondary metabolite, siderophore, and lysine biosynthesis in <italic toggle="yes">aspergillus fumigatus</italic> and is critical for pathogenicity
publisher	American Society for Microbiology
publishDate	2017
url	https://doaj.org/article/cccea62e55644309b3c3b177d1c49935
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A Nonredundant Phosphopantetheinyl Transferase, PptA, Is a Novel Antifungal Target That Directs Secondary Metabolite, Siderophore, and Lysine Biosynthesis in <italic toggle="yes">Aspergillus fumigatus</italic> and Is Critical for Pathogenicity

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