Targeting a critical step in fungal hexosamine biosynthesis

Deborah E. A. Lockhart; Mathew Stanley; Olawale G. Raimi; David A. Robinson; Dominika Boldovjakova; Daniel R. Squair; Andrew T. Ferenbach; Wenxia Fang; Daan M. F. van Aalten

doi:10.1074/jbc.RA120.012985

Targeting a critical step in fungal hexosamine biosynthesis

Deborah E. A. Lockhart^*, Mathew Stanley, Olawale G. Raimi, David A. Robinson, Dominika Boldovjakova, Daniel R. Squair, Andrew T. Ferenbach, Wenxia Fang, Daan M. F. van Aalten^*

^*Corresponding author for this work

University of Dundee

Research output: Contribution to journal › Article › peer-review

12 Citations (Scopus)

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Abstract

Aspergillus fumigatus is a human opportunistic fungal pathogen whose cell wall protects it from the extracellular environment, including host defense responses. Chitin, an essential component of the fungal cell wall, is synthesized from UDP-GlcNAc produced in the hexosamine biosynthetic pathway. Because this pathway is critical for fungal cell wall integrity, the hexosamine biosynthesis enzymes represent potential targets of antifungal drugs. Here, we provide genetic and chemical evidence that glucosamine 6-phosphate N-acetyltransferase (Gna1), a key enzyme in this pathway, is an exploitable antifungal drug target. GNA1 deletion resulted in loss of fungal viability and disruption of the cell wall, phenotypes that could be rescued by exogenous GlcNAc, the product of the Gna1 enzyme. In a murine model of aspergillosis, the Dgna1 mutant strain exhibited attenuated virulence. Using a fragment-based approach, we discovered a small heterocyclic scaffold that binds proximal to the Gna1 active site and can be optimized to a selective sub-micromolar binder. Taken together, we have provided genetic, structural, and chemical evidence that Gna1 is an
antifungal target in A. fumigatus.

Original language	English
Pages (from-to)	8678-8691
Number of pages	14
Journal	Journal of Biological Chemistry
Volume	295
Issue number	26
Early online date	27 Apr 2020
DOIs	https://doi.org/10.1074/jbc.RA120.012985
Publication status	Published - 26 Jun 2020

Bibliographical note

Acknowledgements
We wish to thank the Dundee Drug Discovery Unit for access to the Fragment library and the European Synchrotron Radiation Facility, Grenoble and Diamond Light Source, Oxford for time at the beamline. The assistance from Mr Martin Kierans, School of Life Sciences, University of Dundee with the Electron Microscopy is gratefully acknowledged.

Funding and additional information
This work was supported by a Wellcome Trust Postdoctoral Research Training Fellowship for Clinicians (WT105772/A/14/Z) to DL and an MRC Programme Grant (MR/M004139/1) to DMFvA. DB was funded by a University of Aberdeen Summer Research Scholarship.

Keywords

fungi
Aspergillus fumigatus
cell wall
glucosamine 6-phosphate N-acetyltransferase (Gna1)
resistance
fragment
protein-protein interaction
virulence factor
chitin
antifungal drug development
X-ray crystallography
ASPERGILLUS-FUMIGATUS
PRACTICE GUIDELINES
NIKKOMYCIN-Z
STRESS-RESPONSE
STRUCTURAL VALIDATION
MURINE MODEL
CELL-WALL
GALLERIA-MELLONELLA LARVAE
INHIBITORS
INFECTIOUS-DISEASES SOCIETY

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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Lockhart_et_al_JBC_TargetingACriticalStep_AAM
This research was originally published in the Journal of Biological Chemistry. Author(s). Title. J Biol Chem. Year; Vol:pp-pp. © the Author(s).
Accepted author manuscript, 1.22 MBLicence: Unspecified
J. Biol. Chem.-2020-Lockhart-8678-91_VOR
Author’s Choice—Final version open access under the terms of the Creative Commons CC-BY license https://creativecommons.org/licenses/by/4.0/
Final published version, 1.45 MBLicence: CC BY

Cite this

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title = "Targeting a critical step in fungal hexosamine biosynthesis",

abstract = "Aspergillus fumigatus is a human opportunistic fungal pathogen whose cell wall protects it from the extracellular environment, including host defense responses. Chitin, an essential component of the fungal cell wall, is synthesized from UDP-GlcNAc produced in the hexosamine biosynthetic pathway. Because this pathway is critical for fungal cell wall integrity, the hexosamine biosynthesis enzymes represent potential targets of antifungal drugs. Here, we provide genetic and chemical evidence that glucosamine 6-phosphate N-acetyltransferase (Gna1), a key enzyme in this pathway, is an exploitable antifungal drug target. GNA1 deletion resulted in loss of fungal viability and disruption of the cell wall, phenotypes that could be rescued by exogenous GlcNAc, the product of the Gna1 enzyme. In a murine model of aspergillosis, the Dgna1 mutant strain exhibited attenuated virulence. Using a fragment-based approach, we discovered a small heterocyclic scaffold that binds proximal to the Gna1 active site and can be optimized to a selective sub-micromolar binder. Taken together, we have provided genetic, structural, and chemical evidence that Gna1 is anantifungal target in A. fumigatus.",

keywords = "fungi, Aspergillus fumigatus, cell wall, glucosamine 6-phosphate N-acetyltransferase (Gna1), resistance, fragment, protein-protein interaction, virulence factor, chitin, antifungal drug development, X-ray crystallography, ASPERGILLUS-FUMIGATUS, PRACTICE GUIDELINES, NIKKOMYCIN-Z, STRESS-RESPONSE, STRUCTURAL VALIDATION, MURINE MODEL, CELL-WALL, GALLERIA-MELLONELLA LARVAE, INHIBITORS, INFECTIOUS-DISEASES SOCIETY",

author = "Lockhart, {Deborah E. A.} and Mathew Stanley and Raimi, {Olawale G.} and Robinson, {David A.} and Dominika Boldovjakova and Squair, {Daniel R.} and Ferenbach, {Andrew T.} and Wenxia Fang and {van Aalten}, {Daan M. F.}",

note = "Acknowledgements We wish to thank the Dundee Drug Discovery Unit for access to the Fragment library and the European Synchrotron Radiation Facility, Grenoble and Diamond Light Source, Oxford for time at the beamline. The assistance from Mr Martin Kierans, School of Life Sciences, University of Dundee with the Electron Microscopy is gratefully acknowledged. Funding and additional information This work was supported by a Wellcome Trust Postdoctoral Research Training Fellowship for Clinicians (WT105772/A/14/Z) to DL and an MRC Programme Grant (MR/M004139/1) to DMFvA. DB was funded by a University of Aberdeen Summer Research Scholarship.",

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T1 - Targeting a critical step in fungal hexosamine biosynthesis

AU - Lockhart, Deborah E. A.

AU - Stanley, Mathew

AU - Raimi, Olawale G.

AU - Robinson, David A.

AU - Boldovjakova, Dominika

AU - Squair, Daniel R.

AU - Ferenbach, Andrew T.

AU - Fang, Wenxia

AU - van Aalten, Daan M. F.

N1 - Acknowledgements We wish to thank the Dundee Drug Discovery Unit for access to the Fragment library and the European Synchrotron Radiation Facility, Grenoble and Diamond Light Source, Oxford for time at the beamline. The assistance from Mr Martin Kierans, School of Life Sciences, University of Dundee with the Electron Microscopy is gratefully acknowledged. Funding and additional information This work was supported by a Wellcome Trust Postdoctoral Research Training Fellowship for Clinicians (WT105772/A/14/Z) to DL and an MRC Programme Grant (MR/M004139/1) to DMFvA. DB was funded by a University of Aberdeen Summer Research Scholarship.

PY - 2020/6/26

Y1 - 2020/6/26

N2 - Aspergillus fumigatus is a human opportunistic fungal pathogen whose cell wall protects it from the extracellular environment, including host defense responses. Chitin, an essential component of the fungal cell wall, is synthesized from UDP-GlcNAc produced in the hexosamine biosynthetic pathway. Because this pathway is critical for fungal cell wall integrity, the hexosamine biosynthesis enzymes represent potential targets of antifungal drugs. Here, we provide genetic and chemical evidence that glucosamine 6-phosphate N-acetyltransferase (Gna1), a key enzyme in this pathway, is an exploitable antifungal drug target. GNA1 deletion resulted in loss of fungal viability and disruption of the cell wall, phenotypes that could be rescued by exogenous GlcNAc, the product of the Gna1 enzyme. In a murine model of aspergillosis, the Dgna1 mutant strain exhibited attenuated virulence. Using a fragment-based approach, we discovered a small heterocyclic scaffold that binds proximal to the Gna1 active site and can be optimized to a selective sub-micromolar binder. Taken together, we have provided genetic, structural, and chemical evidence that Gna1 is anantifungal target in A. fumigatus.

AB - Aspergillus fumigatus is a human opportunistic fungal pathogen whose cell wall protects it from the extracellular environment, including host defense responses. Chitin, an essential component of the fungal cell wall, is synthesized from UDP-GlcNAc produced in the hexosamine biosynthetic pathway. Because this pathway is critical for fungal cell wall integrity, the hexosamine biosynthesis enzymes represent potential targets of antifungal drugs. Here, we provide genetic and chemical evidence that glucosamine 6-phosphate N-acetyltransferase (Gna1), a key enzyme in this pathway, is an exploitable antifungal drug target. GNA1 deletion resulted in loss of fungal viability and disruption of the cell wall, phenotypes that could be rescued by exogenous GlcNAc, the product of the Gna1 enzyme. In a murine model of aspergillosis, the Dgna1 mutant strain exhibited attenuated virulence. Using a fragment-based approach, we discovered a small heterocyclic scaffold that binds proximal to the Gna1 active site and can be optimized to a selective sub-micromolar binder. Taken together, we have provided genetic, structural, and chemical evidence that Gna1 is anantifungal target in A. fumigatus.

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KW - virulence factor

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KW - STRESS-RESPONSE

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KW - MURINE MODEL

KW - CELL-WALL

KW - GALLERIA-MELLONELLA LARVAE

KW - INHIBITORS

KW - INFECTIOUS-DISEASES SOCIETY

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U2 - 10.1074/jbc.RA120.012985

DO - 10.1074/jbc.RA120.012985

M3 - Article

C2 - 32341126

SN - 0021-9258

VL - 295

SP - 8678

EP - 8691

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

IS - 26

ER -

Targeting a critical step in fungal hexosamine biosynthesis

Abstract

Bibliographical note

Keywords

UN SDGs

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