Data Availability: The authors confirm that all data underlying the findings are fully available without restriction. Raw data have been deposited in the Gene Expression Omnibus (GEO) (http://www.ncbi.nlm.nih.gov/geo/) under accession number GSE54810. Funding: This work was supported in part by grants to EMB from the MRC (G0501164) and BBSRC (BB/G009619/1), to EMB and NDR from the Wellcome Trust (WT093596MA), to MB from Imperial College London (Division of Investigative Sciences PhD Studentship), to HH from the ERA-NET PathoGenoMics project TRANSPAT, Austrian Science Foundation (FWF I282-B09), to SGF from the National Institutes of Health, USA (R01AI073829). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Peer reviewedPublisher PD

Alcazar-Fuoli, Laura

Armstrong-James, Darius

Bertuzzi, Margherita

Bignell, Elaine M

Cairns, Timothy C

Chen, Dan

Cheverton, Angela M

Espeso, Eduardo A

Fedorova, Natalie D

Filler, Scott G

Haas, Hubertus

Herbst, Susanne

Liu, Hong

Munro, Carol A

Muñoz, Alberto

Nierman, William C

Read, Nick D

Safari, Maryam

Saijo, Shinobu

Schrettl, Markus

Walker, Louise A

PLoS Pathogens

English

PubMed

Margherita Bertuzzi

Markus Schrettl

Laura Alcazar-Fuoli

Timothy C. Cairns

Alberto Muñoz

Louise A. Walker

Susanne Herbst

Maryam Safari

Angela M. Cheverton

Dan Chen

Hong Liu

Shinobu Saijo

Natalie D. Fedorova

Darius Armstrong-James

Carol A. Munro

Nick D. Read

Scott G. Filler

Eduardo A. Espeso

William C. Nierman

Hubertus Haas

Elaine M. Bignell

Crossref

Correction: The pH-Responsive PacC Transcription Factor of Aspergillus fumigatus Governs Epithelial Entry and Tissue Invasion during Pulmonary Aspergillosis

Correction: The pH-Responsive PacC Transcription Factor of Aspergillus fumigatus Governs Epithelial Entry and Tissue Invasion during Pulmonary Aspergillosis. PLoS Pathog. 2015 Jun 18;11(6):e1004943. doi: 10.1371/journal.ppat.1004943. PMID: 26086393.Correction: The pH-responsive PacC transcription factor of Aspergillus fumigatus governs epithelial entry and tissue invasion during pulmonary aspergillosis. PLoS Pathog. 2015 Apr 8;11(4):e1004802. doi: 10.1371/journal.ppat.1004802. PMID: 25853727.Destruction of the pulmonary epithelium is a major feature of lung diseases caused by the mould pathogen Aspergillus fumigatus. Although it is widely postulated that tissue invasion is governed by fungal proteases, A. fumigatus mutants lacking individual or multiple enzymes remain fully invasive, suggesting a concomitant requirement for other pathogenic activities during host invasion. In this study we discovered, and exploited, a novel, tissue non-invasive, phenotype in A. fumigatus mutants lacking the pH-responsive transcription factor PacC. Our study revealed a novel mode of epithelial entry, occurring in a cell wall-dependent manner prior to protease production, and via the Dectin-1 β-glucan receptor. ΔpacC mutants are defective in both contact-mediated epithelial entry and protease expression, and significantly attenuated for pathogenicity in leukopenic mice. We combined murine infection modelling, in vivo transcriptomics, and in vitro infections of human alveolar epithelia, to delineate two major, and sequentially acting, PacC-dependent processes impacting epithelial integrity in vitro and tissue invasion in the whole animal. We demonstrate that A. fumigatus spores and germlings are internalised by epithelial cells in a contact-, actin-, cell wall- and Dectin-1 dependent manner and ΔpacC mutants, which aberrantly remodel the cell wall during germinative growth, are unable to gain entry into epithelial cells, both in vitro and in vivo. We further show that PacC acts as a global transcriptional regulator of secreted molecules during growth in the leukopenic mammalian lung, and profile the full cohort of secreted gene products expressed during invasive infection. Our study reveals a combinatorial mode of tissue entry dependent upon sequential, and mechanistically distinct, perturbations of the pulmonary epithelium and demonstrates, for the first time a protective role for Dectin-1 blockade in epithelial defences. Infecting ΔpacC mutants are hypersensitive to cell wall-active antifungal agents highlighting the value of PacC signalling as a target for antifungal therapy.This work was supported in part by grants to EMB from the MRC (G0501164) and BBSRC (BB/G009619/1), to EMB and NDR from the Wellcome Trust (WT093596MA), to MB from Imperial College London (Division of Investigative Sciences PhD Studentship), to HH from the ERA-NET PathoGenoMics project TRANSPAT, Austrian Science Foundation (FWF I282-B09), to SGF from the National Institutes of Health, USA (R01AI073829). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.S

REPISALUD

The pH-responsive PacC transcription factor of Aspergillus fumigatus governs epithelial entry and tissue invasion during pulmonary aspergillosis

Data Availability: The authors confirm that all data underlying the findings are fully available without restriction. Raw data have been deposited in the Gene Expression Omnibus (GEO) (http://www.ncbi.nlm.nih.gov/geo/) under accession number GSE54810. Funding: This work was supported in part by grants to EMB from the MRC (G0501164) and BBSRC (BB/G009619/1), to EMB and NDR from the Wellcome Trust (WT093596MA), to MB from Imperial College London (Division of Investigative Sciences PhD Studentship), to HH from the ERA-NET PathoGenoMics project TRANSPAT, Austrian Science Foundation (FWF I282-B09), to SGF from the National Institutes of Health, USA (R01AI073829). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Peer reviewe

Aberdeen University Research

eScholarship - University of California

Bertuzzi, M

Schrettl, M

Alcazar-Fuoli, L

Cairns, TC

Muñoz, A

Walker, LA

Herbst, S

Safari, M

Cheverton, AM

Chen, D

Liu, H

Saijo, S

Fedorova, ND

Armstrong-James, D

Munro, CA

Read, ND

Filler, SG

Espeso, EA

Nierman, WC

Haas, H

Bignell, EM

Spiral - Imperial College Digital Repository

CORRECTIONCorrection: The pH-Responsive PacCTranscription Factor of Aspergillus fumigatusGoverns Epithelial Entry and Tissue Invasionduring Pulmonary AspergillosisMargherita Bertuzzi, Markus Schrettl, Laura Alcazar-Fuoli, Timothy C. Cairns,Alberto Muñoz, Louise A. Walker, Susanne Herbst, Maryam Safari, Angela M. Cheverton,Dan Chen, Hong Liu, Shinobu Saijo, Natalie D. Fedorova, Darius Armstrong-James, CarolA. Munro, Nick D. Read, Scott G. Filler, Eduardo A. Espeso, William C. Nierman,Hubertus Haas, Elaine M. BignellThe authors would like to correct the following sentence in the Materials and Methods sectionof the article:“For preparation of A. fumigatus culture supernatants, 106 spores/ml were grown in Dul-becco’s Modified Eagle Medium (DMEM, Sigma) supplemented with 10% foetal bovine serum(FBS, Sigma) and 10% penicillin and streptomycin (Sigma) at 37uC, 5% CO2 for either 16, 48,60 or 72 hr.”In this study, Dulbecco's Modified Eagle Medium (DMEM, Sigma) was supplemented with10% foetal bovine serum (FBS, Sigma) and 1% penicillin and streptomycin (Sigma), not 10%penicillin and streptomycin (Sigma) as originally indicated.References1. Bertuzzi M, Schrettl M, Alcazar-Fuoli L, Cairns TC, Muñoz A, Walker LA, et al. (2014) The pH-Respon-sive PacC Transcription Factor of Aspergillus fumigatusGoverns Epithelial Entry and Tissue Invasionduring Pulmonary Aspergillosis. PLoS Pathog 10(10): e1004413. doi:10.1371/journal.ppat.1004413PMID: 25329394PLOS Pathogens | DOI:10.1371/journal.ppat.1004943 June 18, 2015 1 / 1OPEN ACCESSCitation: Bertuzzi M, Schrettl M, Alcazar-Fuoli L,Cairns TC, Muñoz A, Walker LA, et al. (2015)Correction: The pH-Responsive PacC TranscriptionFactor of Aspergillus fumigatus Governs EpithelialEntry and Tissue Invasion during PulmonaryAspergillosis. PLoS Pathog 11(6): e1004943.doi:10.1371/journal.ppat.1004943Published: June 18, 2015Copyright: © 2015 Bertuzzi et al. This is an openaccess article distributed under the terms of theCreative Commons Attribution License, which permitsunrestricted use, distribution, and reproduction in anymedium, provided the original author and source arecredited.

Correction: The pH-Responsive PacC Transcription Factor of Aspergillus fumigatus Governs Epithelial Entry and Tissue Invasion during Pulmonary Aspergillosis.

Destruction of the pulmonary epithelium is a major feature of lung diseases caused by the mould pathogen Aspergillus fumigatus. Although it is widely postulated that tissue invasion is governed by fungal proteases, A. fumigatus mutants lacking individual or multiple enzymes remain fully invasive, suggesting a concomitant requirement for other pathogenic activities during host invasion. In this study we discovered, and exploited, a novel, tissue non-invasive, phenotype in A. fumigatus mutants lacking the pH-responsive transcription factor PacC. Our study revealed a novel mode of epithelial entry, occurring in a cell wall-dependent manner prior to protease production, and via the Dectin-1 β-glucan receptor. ΔpacC mutants are defective in both contact-mediated epithelial entry and protease expression, and significantly attenuated for pathogenicity in leukopenic mice. We combined murine infection modelling, in vivo transcriptomics, and in vitro infections of human alveolar epithelia, to delineate two major, and sequentially acting, PacC-dependent processes impacting epithelial integrity in vitro and tissue invasion in the whole animal. We demonstrate that A. fumigatus spores and germlings are internalised by epithelial cells in a contact-, actin-, cell wall- and Dectin-1 dependent manner and ΔpacC mutants, which aberrantly remodel the cell wall during germinative growth, are unable to gain entry into epithelial cells, both in vitro and in vivo. We further show that PacC acts as a global transcriptional regulator of secreted molecules during growth in the leukopenic mammalian lung, and profile the full cohort of secreted gene products expressed during invasive infection. Our study reveals a combinatorial mode of tissue entry dependent upon sequential, and mechanistically distinct, perturbations of the pulmonary epithelium and demonstrates, for the first time a protective role for Dectin-1 blockade in epithelial defences. Infecting ΔpacC mutants are hypersensitive to cell wall-active antifungal agents highlighting the value of PacC signalling as a target for antifungal therapy

Timothy C Cairns

Louise A Walker

Angela M Cheverton

Natalie D Fedorova

Carol A Munro

Nick D Read

Scott G Filler

Eduardo A Espeso

William C Nierman

Elaine M Bignell

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The pH-responsive PacC transcription factor of Aspergillus fumigatus governs epithelial entry and tissue invasion during pulmonary aspergillosis.

The University of Manchester - Institutional Repository

https://repisalud.isciii.es/bitstream/20.500.12105/6837/1/ThePH-ResponsivePacCTranscription_2014.pdf

The pH-responsive PacC transcription factor of Aspergillus fumigatus governs epithelial entry and tissue invasion during pulmonary aspergillosis

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Aberdeen University Research

eScholarship - University of California

Spiral - Imperial College Digital Repository

Directory of Open Access Journals

The University of Manchester - Institutional Repository

Directory of Open Access Journals