{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,3]],"date-time":"2026-01-03T21:07:33Z","timestamp":1767474453450,"version":"build-2065373602"},"reference-count":55,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2022,7,16]],"date-time":"2022-07-16T00:00:00Z","timestamp":1657929600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000265","name":"Medical Research Council Centre for Medical Mycology at the University of Exeter","doi-asserted-by":"publisher","award":["MR\/N006364\/2","MR\/M026663\/2","NNF20OC0059614"],"award-info":[{"award-number":["MR\/N006364\/2","MR\/M026663\/2","NNF20OC0059614"]}],"id":[{"id":"10.13039\/501100000265","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000265","name":"Medical Research Council","doi-asserted-by":"publisher","award":["MR\/N006364\/2","MR\/M026663\/2","NNF20OC0059614"],"award-info":[{"award-number":["MR\/N006364\/2","MR\/M026663\/2","NNF20OC0059614"]}],"id":[{"id":"10.13039\/501100000265","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Ben Boyd\u2019d Novo Nordisk Foundation Laureate Research Fellowship","award":["MR\/N006364\/2","MR\/M026663\/2","NNF20OC0059614"],"award-info":[{"award-number":["MR\/N006364\/2","MR\/M026663\/2","NNF20OC0059614"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["JoF"],"abstract":"<jats:p>Aspergillus fumigatus is the leading cause of the fungal invasive disease called aspergillosis, which is associated with a high mortality rate that can reach 50% in some groups of immunocompromised individuals. The increasing prevalence of azole-resistant A. fumigatus isolates, both in clinical settings and the environment, highlights the importance of discovering new fungal virulence factors that can potentially become targets for novel antifungals. Nitronate monooxygenases (Nmos) represent potential targets for antifungal compounds as no orthologs of those enzymes are present in humans. Nmos catalyse the denitrification of nitroalkanes, thereby detoxifying these mediators of nitro-oxidative stress, and therefore we tested whether Nmos provide protection for A. fumigatus against host-imposed stresses at sites of infection. The results of inhibition zone assays indicated that Nmo2 and Nmo5 are not essential for the oxidative stress resistance of A. fumigatus in vitro. In addition, the resazurin-based metabolic activity assay revealed that the growth of mutants lacking the nmo2 or nmo5 genes was only slightly reduced in the presence of 0.05 mM peroxynitrite. Nevertheless, both Nmo2 and Nmo5 were shown to contribute to defense against murine bone marrow-derived macrophages, and this was no longer observed when NADPH oxidase, the main generator of reactive oxygen species during infection, was inhibited in macrophages. Furthermore, we revealed that Nnmos promote the virulence of the fungus in the Galleria mellonella model of infection. Both nmo2 and nmo5 knock-out strains were less virulent than the wild-type control as recorded 72 h post-infection. Our results indicate that Nmos play a role in the virulence of A. fumigatus.<\/jats:p>","DOI":"10.3390\/jof8070736","type":"journal-article","created":{"date-parts":[[2022,7,17]],"date-time":"2022-07-17T22:57:06Z","timestamp":1658098626000},"page":"736","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Understanding the Role of Nitronate Monooxygenases in Virulence of the Human Fungal Pathogen Aspergillus fumigatus"],"prefix":"10.3390","volume":"8","author":[{"given":"Phuong Tuyen","family":"Nguyen","sequence":"first","affiliation":[{"name":"MRC Centre for Medical Mycology, University of Exeter, Geoffrey Pope Building, Stocker Road, Exeter EX4 4QD, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1789-2346","authenticated-orcid":false,"given":"Theresa","family":"Wacker","sequence":"additional","affiliation":[{"name":"MRC Centre for Medical Mycology, University of Exeter, Geoffrey Pope Building, Stocker Road, Exeter EX4 4QD, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1406-4251","authenticated-orcid":false,"given":"Alistair J. P.","family":"Brown","sequence":"additional","affiliation":[{"name":"MRC Centre for Medical Mycology, University of Exeter, Geoffrey Pope Building, Stocker Road, Exeter EX4 4QD, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alessandra","family":"da Silva Dantas","sequence":"additional","affiliation":[{"name":"MRC Centre for Medical Mycology, University of Exeter, Geoffrey Pope Building, Stocker Road, Exeter EX4 4QD, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3747-1856","authenticated-orcid":false,"given":"Elena","family":"Shekhova","sequence":"additional","affiliation":[{"name":"MRC Centre for Medical Mycology, University of Exeter, Geoffrey Pope Building, Stocker Road, Exeter EX4 4QD, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,7,16]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Bongomin, F., Gago, S., Oladele, R.O., and Denning, D.W. (2017). Global and Multi-National Prevalence of Fungal Diseases-Estimate Precision. J. Fungi, 3.","DOI":"10.3390\/jof3040057"},{"key":"ref_2","unstructured":"WHO, G. (2020). Global Tuberculosis Report 2020. Glob. Tuberc. Rep., 2020, Available online: https:\/\/www.who.int\/publications\/i\/item\/9789240013131."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"e00140-18","DOI":"10.1128\/CMR.00140-18","article-title":"Aspergillus fumigatus and Aspergillosis in 2019","volume":"33","author":"Chamilos","year":"2019","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_4","first-page":"1068","article-title":"The epidemiology of fungal infections in patients with hematologic malignancies: The SEIFEM-2004 study","volume":"91","author":"Pagano","year":"2006","journal-title":"Haematologica"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"S49","DOI":"10.1080\/13693780400020113","article-title":"Incidence of invasive aspergillosis following hematopoietic stem cell and solid organ transplantation: Interim results of a prospective multicenter surveillance program","volume":"43","author":"Morgan","year":"2005","journal-title":"Med. Mycol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"135","DOI":"10.1111\/1348-0421.12957","article-title":"Azole-resistant Aspergillus fumigatus as an emerging worldwide pathogen","volume":"66","author":"Zea","year":"2022","journal-title":"Microbiol. Immunol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"739","DOI":"10.1126\/science.aap7999","article-title":"Worldwide emergence of resistance to antifungal drugs challenges human health and food security","volume":"360","author":"Fisher","year":"2018","journal-title":"Science"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"7632","DOI":"10.1111\/1462-2920.15660","article-title":"Azole-resistant Aspergillus fumigatus is highly prevalent in the environment of Vietnam, with marked variability by land use type","volume":"23","author":"Duong","year":"2021","journal-title":"Environ. Microbiol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"jkab427","DOI":"10.1093\/g3journal\/jkab427","article-title":"Evidence for the agricultural origin of resistance to multiple antimicrobials in Aspergillus fumigatus, a fungal pathogen of humans","volume":"12","author":"Kang","year":"2022","journal-title":"G3"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"34","DOI":"10.1016\/j.semcdb.2018.03.004","article-title":"Oxidative responses and fungal infection biology","volume":"89","author":"Warris","year":"2019","journal-title":"Semin. Cell Dev. Biol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"835","DOI":"10.1128\/EC.3.4.835-846.2004","article-title":"Mechanisms of resistance to oxidative and nitrosative stress: Implications for fungal survival in mammalian hosts","volume":"3","author":"Missall","year":"2004","journal-title":"Eukaryot. Cell"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"17054","DOI":"10.1038\/nmicrobiol.2017.54","article-title":"The Magnaporthe oryzae nitrooxidative stress response suppresses rice innate immunity during blast disease","volume":"2","author":"Hartline","year":"2017","journal-title":"Nat. Microbiol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"378","DOI":"10.1007\/s002940050350","article-title":"Development of a homologous transformation system for the human pathogenic fungus Aspergillus fumigatus based on the pyrG gene encoding orotidine 5\u2032-monophosphate decarboxylase","volume":"33","author":"Weidner","year":"1998","journal-title":"Curr. Genet."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"76","DOI":"10.1111\/j.1365-2958.2006.05215.x","article-title":"The Aspergillus nidulans F-box protein GrrA links SCF activity to meiosis","volume":"61","author":"Krappmann","year":"2006","journal-title":"Mol. Microbiol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1016\/S0065-2660(08)60408-3","article-title":"The genetics of Aspergillus nidulans","volume":"5","author":"Pontecorvo","year":"1953","journal-title":"Adv. Genet."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"237","DOI":"10.1007\/s004380050494","article-title":"The Aspergillus nidulans lysF gene encodes homoaconitase, an enzyme involved in the fungus-specific lysine biosynthesis pathway","volume":"255","author":"Weidner","year":"1997","journal-title":"Mol. Gen. Genet."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"641495","DOI":"10.3389\/fimmu.2021.641495","article-title":"Mitochondrial Reactive Oxygen Species Regulate Immune Responses of Macrophages to Aspergillus fumigatus","volume":"12","author":"Hatinguais","year":"2021","journal-title":"Front. Immunol."},{"key":"ref_18","unstructured":"Davies, J.Q., and Gordon, S. (2005). Isolation and culture of murine macrophages. Basic Cell Culture Protocols, Springer."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"64","DOI":"10.1038\/nprot.2013.161","article-title":"Resin-assisted enrichment of thiols as a general strategy for proteomic profiling of cysteine-based reversible modifications","volume":"9","author":"Guo","year":"2014","journal-title":"Nat. Protoc."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"2987","DOI":"10.1007\/s00253-020-10450-0","article-title":"Members of the nitronate monooxygenase gene family from Metarhizium brunneum are induced during the process of infection to Plutella xylostella","volume":"104","year":"2020","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"315","DOI":"10.1152\/physrev.00029.2006","article-title":"Nitric oxide and peroxynitrite in health and disease","volume":"87","author":"Pacher","year":"2007","journal-title":"Physiol. Rev."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1111\/mmi.13211","article-title":"Nitric oxide synthesis by nitrate reductase is regulated during development in Aspergillus","volume":"99","author":"Marcos","year":"2016","journal-title":"Mol. Microbiol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"542","DOI":"10.1177\/1087057111433459","article-title":"A new approach to drug discovery: High-throughput screening of microbial natural extracts against Aspergillus fumigatus using resazurin","volume":"17","author":"Monteiro","year":"2012","journal-title":"J. Biomol. Screen."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1016\/0891-5849(94)00105-S","article-title":"A practical method for preparing peroxynitrite solutions of low ionic strength and free of hydrogen peroxide","volume":"18","author":"Pryor","year":"1995","journal-title":"Free. Radic. Biol. Med."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"4809","DOI":"10.2741\/3569","article-title":"Role of peroxynitrite in the redox regulation of cell signal transduction pathways","volume":"14","author":"Liaudet","year":"2009","journal-title":"Front. Biosci. (Landmark Ed)"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"111","DOI":"10.1042\/bj2370111","article-title":"The effect of the inhibitor diphenylene iodonium on the superoxide-generating system of neutrophils. Specific labelling of a component polypeptide of the oxidase","volume":"237","author":"Cross","year":"1986","journal-title":"Biochem. J."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1016\/j.chom.2015.12.002","article-title":"Aspergillus Cell Wall Melanin Blocks LC3-Associated Phagocytosis to Promote Pathogenicity","volume":"19","author":"Akoumianaki","year":"2016","journal-title":"Cell Host Microbe"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"e93156","DOI":"10.1172\/jci.insight.93156","article-title":"Immunity against fungi","volume":"2","author":"Lionakis","year":"2017","journal-title":"JCI Insight"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"240","DOI":"10.1007\/s10886-016-0684-0","article-title":"Two Defensive Lines in Juvenile Leaf Beetles; Esters of 3-nitropropionic Acid in the Hemolymph and Aposematic Warning","volume":"42","author":"Pauls","year":"2016","journal-title":"J. Chem. Ecol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1787","DOI":"10.1007\/s10886-007-9341-y","article-title":"Nitroalkenes and sesquiterpene hydrocarbons from the frontal gland of three prorhinotermes termite species","volume":"33","author":"Piskorski","year":"2007","journal-title":"J. Chem. Ecol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"8019","DOI":"10.1007\/s00253-021-11623-1","article-title":"Peculiarities of nitronate monooxygenases and perspectives for in vivo and in vitro applications","volume":"105","year":"2021","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"84","DOI":"10.1016\/j.abb.2012.03.015","article-title":"A novel activity for fungal nitronate monooxygenase: Detoxification of the metabolic inhibitor propionate-3-nitronate","volume":"521","author":"Francis","year":"2012","journal-title":"Arch. Biochem. Biophys."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"3551","DOI":"10.1128\/IAI.71.6.3551-3562.2003","article-title":"Catalases of Aspergillus fumigatus","volume":"71","author":"Paris","year":"2003","journal-title":"Infect. Immun."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"568","DOI":"10.1094\/MPMI-20-5-0568","article-title":"A novel role for catalase B in the maintenance of fungal cell-wall integrity during host invasion in the rice blast fungus Magnaporthe grisea","volume":"20","author":"Skamnioti","year":"2007","journal-title":"Mol. Plant Microbe Interact."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1094\/MPMI-07-10-0175","article-title":"The role of catalase-peroxidase secreted by Magnaporthe oryzae during early infection of rice cells","volume":"24","author":"Tanabe","year":"2011","journal-title":"Mol. Plant Microbe Interact."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"119","DOI":"10.1002\/jbt.20061","article-title":"Response of the antioxidant defense system to tert-butyl hydroperoxide and hydrogen peroxide in a human hepatoma cell line (HepG2)","volume":"19","author":"Ramos","year":"2005","journal-title":"J. Biochem. Mol. Toxicol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"682","DOI":"10.1016\/j.fgb.2007.01.009","article-title":"Characterization of the SKN7 ortholog of Aspergillus fumigatus","volume":"44","author":"Lamarre","year":"2007","journal-title":"Fungal Genet. Biol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"2290","DOI":"10.1128\/EC.00267-07","article-title":"The Aspergillus fumigatus transcriptional regulator AfYap1 represents the major regulator for defense against reactive oxygen intermediates but is dispensable for pathogenicity in an intranasal mouse infection model","volume":"6","author":"Lessing","year":"2007","journal-title":"Eukaryot. Cell"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"30667","DOI":"10.1074\/jbc.M705929200","article-title":"NOS2 regulation of NF-kappaB by S-nitrosylation of p65","volume":"282","author":"Kelleher","year":"2007","journal-title":"J. Biol. Chem."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1814","DOI":"10.1038\/srep01814","article-title":"S-nitrosylation of ERK inhibits ERK phosphorylation and induces apoptosis","volume":"3","author":"Feng","year":"2013","journal-title":"Sci. Rep."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1525","DOI":"10.1093\/infdis\/jit049","article-title":"Rv1894c is a novel hypoxia-induced nitronate monooxygenase required for Mycobacterium tuberculosis virulence","volume":"207","author":"Klinkenberg","year":"2013","journal-title":"J. Infect. Dis."},{"key":"ref_42","first-page":"1","article-title":"Molecular modeling and docking study of 2-nitropropane dioxygenase of Mycobacterium tuberculosis","volume":"6","author":"Abid","year":"2017","journal-title":"Int. J. Biomed. Data Min."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"ftab013","DOI":"10.1093\/femspd\/ftab013","article-title":"Fungal immunity and pathogenesis in mammals versus the invertebrate model organism Galleria mellonella","volume":"79","author":"Casadevall","year":"2021","journal-title":"Pathog. Dis."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"785","DOI":"10.1038\/nmeth.1701","article-title":"SignalP 4.0: Discriminating Signal Peptides from Transmembrane Regions","volume":"8","author":"Petersen","year":"2011","journal-title":"Nat. Methods"},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"Blum, T., Briesemeister, S., and Kohlbacher, O. (2009). MultiLoc2: Integrating Phylogeny and Gene Ontology Terms Improves Subcellular Protein Localization Prediction. BMC Bioinform., 10.","DOI":"10.1186\/1471-2105-10-274"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"5363","DOI":"10.1021\/pr900665y","article-title":"SherLoc2: A High-Accuracy Hybrid Method for Predicting Subcellular Localization of Proteins","volume":"8","author":"Briesemeister","year":"2009","journal-title":"J. Proteome Res."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"W497","DOI":"10.1093\/nar\/gkq477","article-title":"YLoc\u2014an Interpretable Web Server for Predicting Subcellular Localization","volume":"38","author":"Briesemeister","year":"2010","journal-title":"Nucleic Acids Res."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"19448","DOI":"10.1038\/s41598-019-55351-z","article-title":"OutCyte: A Novel Tool for Predicting Unconventional Protein Secretion","volume":"9","author":"Zhao","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_49","doi-asserted-by":"crossref","unstructured":"Camacho, C., Coulouris, G., Avagyan, V., Ma, N., Papadopoulos, J., Bealer, K., and Madden, T.L. (2009). BLAST+: Architecture and Applications. BMC Bioinform., 10.","DOI":"10.1186\/1471-2105-10-421"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"D265","DOI":"10.1093\/nar\/gkz991","article-title":"CDD\/SPARCLE: The Conserved Domain Database in 2020","volume":"48","author":"Lu","year":"2020","journal-title":"Nucleic Acids Res."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"D344","DOI":"10.1093\/nar\/gkaa977","article-title":"The InterPro Protein Families and Domains Database: 20 Years On","volume":"49","author":"Blum","year":"2021","journal-title":"Nucleic Acids Res."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"772","DOI":"10.1093\/molbev\/mst010","article-title":"MAFFT Multiple Sequence Alignment Software Version 7: Improvements in Performance and Usability","volume":"30","author":"Katoh","year":"2013","journal-title":"Mol. Biol. Evol."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"268","DOI":"10.1093\/molbev\/msu300","article-title":"IQ-TREE: A Fast and Effective Stochastic Algorithm for Estimating Maximum-Likelihood Phylogenies","volume":"32","author":"Nguyen","year":"2015","journal-title":"Mol. Biol. Evol."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"587","DOI":"10.1038\/nmeth.4285","article-title":"ModelFinder: Fast Model Selection for Accurate Phylogenetic Estimates","volume":"14","author":"Kalyaanamoorthy","year":"2017","journal-title":"Nat. Methods"},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"4380","DOI":"10.1128\/iai.62.10.4380-4388.1994","article-title":"Rodletless Mutants of Aspergillus Fumigatus","volume":"62","author":"Thau","year":"1994","journal-title":"Infect. Immun"}],"container-title":["Journal of Fungi"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2309-608X\/8\/7\/736\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T23:52:03Z","timestamp":1760140323000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2309-608X\/8\/7\/736"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,7,16]]},"references-count":55,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2022,7]]}},"alternative-id":["jof8070736"],"URL":"https:\/\/doi.org\/10.3390\/jof8070736","relation":{},"ISSN":["2309-608X"],"issn-type":[{"type":"electronic","value":"2309-608X"}],"subject":[],"published":{"date-parts":[[2022,7,16]]}}}