{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T16:15:36Z","timestamp":1791389736708,"version":"4.3.3"},"reference-count":68,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,7,25]],"date-time":"2025-07-25T00:00:00Z","timestamp":1753401600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:sec><jats:title>Introduction<\/jats:title><jats:p>The ability to acquire iron and maintain iron homeostasis is crucial for the virulence of the human pathogenic fungus <jats:italic>Cryptococcus neoformans<\/jats:italic>. This study investigates the role of Bud32, a core virulence kinase and component of the KEOPS complex, within the iron regulatory network of <jats:italic>C. neoformans.<\/jats:italic><\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>We used gene deletion techniques to study the phenotypic effects of <jats:italic>BUD32<\/jats:italic> gene knockout and conducted proteomic and metabolomic analyses to assess changes in protein expression and metabolite levels in the mutant. Additionally, we performed <jats:italic>in vivo<\/jats:italic> phosphoproteomics analysis to evaluate Bud32 impact on iron regulatory proteins.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Our findings revealed that deletion of <jats:italic>BUD32<\/jats:italic> gene significantly impaired growth in iron-limiting environments, leading to notable alterations in the expression of iron transport and iron-sulfur cluster (ISC)-containing proteins. Specifically, Bud32 was shown to modulate ISC assembly and influence the activity of key iron-sulfur binding proteins, including Grx4, Cir1, and HapX. Metabolic profiling indicated changes in 696 metabolites, with reductions in biliverdin levels. Additionally, <jats:italic>BUD32<\/jats:italic> gene deletion resulted in widespread changes in the phosphorylation status of numerous proteins, including the iron regulators Cir1 and Rim101.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>These findings provide evidence for the involvement of the kinase Bud32 in regulating iron homeostasis in <jats:italic>C. neoformans<\/jats:italic>, thereby contributing to our understanding of its virulence mechanisms.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fimmu.2025.1624237","type":"journal-article","created":{"date-parts":[[2025,7,25]],"date-time":"2025-07-25T11:19:37Z","timestamp":1753442377000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":3,"title":["The kinase Bud32 regulates iron homeostasis in fungal pathogen Cryptococcus neoformans"],"prefix":"10.3389","volume":"16","author":[{"given":"Yuanyuan","family":"Ma","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bo","family":"Pan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenzhi","family":"Lei","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenjie","family":"Fang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Weihua","family":"Pan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wanqing","family":"Liao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bin","family":"Xu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peng","family":"Xue","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2025,7,25]]},"reference":[{"key":"B1","first-page":"647","article-title":"Tackling human fungal infections","volume-title":"Science","author":"Brown","year":"2012"},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.1038\/s41579-022-00720-1","article-title":"Tackling the emerging threat of antifungal resistance to human health","volume":"20","author":"Fisher","year":"2022","journal-title":"Nat Rev Microbiol"},{"key":"B3","doi-asserted-by":"publisher","DOI":"10.1038\/s41579-023-00960-9","article-title":"The rapid emergence of antifungal-resistant human-pathogenic fungi","volume":"1","author":"Lockhart","year":"2023","journal-title":"Nat Rev Microbiol"},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.7573\/dic.2023-9-2","article-title":"Invasive fungal infections in critically ill children: epidemiology, risk factors and antifungal drugs","volume":"13","author":"Hon","year":"2024","journal-title":"Drugs Context"},{"key":"B5","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1080\/21501203.2023.2295409","article-title":"The pathways and the mechanisms by which Cryptococcus enters the brain","author":"Zhou","year":"2024","journal-title":"Mycology"},{"key":"B6","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1080\/21501203.2023.2285764","article-title":"Progress in the application of nanoparticles for the treatment of fungal infections: A review","volume":"15","author":"Zhu","year":"2024","journal-title":"Mycology"},{"key":"B7","volume-title":"WHO fungal priority pathogens list to guide research, development and public health action","author":"Organization","year":"2022"},{"key":"B8","doi-asserted-by":"publisher","DOI":"10.1080\/21501203.2023.2295409","article-title":"The pathways and the mechanisms by which Cryptococcus enters the brain","volume":"15","author":"Zhou","year":"2024","journal-title":"Mycology"},{"key":"B9","doi-asserted-by":"publisher","DOI":"10.1016\/S1473-3099(17)30243-8","article-title":"Global burden of disease of HIV-associated cryptococcal meningitis: an updated analysis","volume":"17","author":"Rajasingham","year":"2017","journal-title":"Lancet Infect Dis"},{"key":"B10","doi-asserted-by":"publisher","DOI":"10.1038\/s41579-021-00511-0","article-title":"Treatment strategies for cryptococcal infection: challenges, advances and future outlook","volume":"19","author":"Iyer","year":"2021","journal-title":"Nat Rev Microbiol"},{"key":"B11","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2025.1610743","article-title":"AI in fungal drug development: opportunities, challenges, and future outlook","volume":"15","author":"Li","year":"2025","journal-title":"Front Cell Infection Microbiol"},{"key":"B12","doi-asserted-by":"publisher","DOI":"10.1080\/21501203.2025.2473507","article-title":"Deciphering the role of mitochondria in human fungal drug resistance","author":"Ma","year":"2025","journal-title":"Mycology"},{"key":"B13","article-title":"The role of lactoferrin in the iron metabolism. Part II. Antimicrobial and antiinflammatory effect of lactoferrin by chelation of iron","volume":"64","author":"Artym","year":"2010","journal-title":"Postepy Hig Med Dosw (Online)"},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.3389\/fpara.2023.1330398","article-title":"Antiparasitic activity of the iron-containing milk protein lactoferrin and its potential derivatives against human intestinal and blood parasites","volume":"2","author":"Anand","year":"2023","journal-title":"Front Parasitol"},{"key":"B15","doi-asserted-by":"publisher","first-page":"346","DOI":"10.3201\/eid0503.990305","article-title":"Iron loading and disease surveillance","volume":"5","author":"Weinberg","year":"1999","journal-title":"Emerging Infect Dis"},{"key":"B16","doi-asserted-by":"publisher","DOI":"10.1038\/nrmicro1046","article-title":"Iron and microbial infection","volume":"2","author":"Schaible","year":"2004","journal-title":"Nat Rev Microbiol"},{"key":"B17","doi-asserted-by":"publisher","DOI":"10.1126\/science.1224577","article-title":"Hepcidin and the iron-infection axis","volume":"338","author":"Drakesmith","year":"2012","journal-title":"science"},{"key":"B18","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2013.00080","article-title":"Shared and distinct mechanisms of iron acquisition by bacterial and fungal pathogens of humans","volume":"3","author":"Caza","year":"2013","journal-title":"Front Cell infection Microbiol"},{"key":"B19","doi-asserted-by":"publisher","DOI":"10.1093\/infdis\/167.1.186","article-title":"Regulation of cryptococcal capsular polysaccharide by iron","volume":"167","author":"Vartivarian","year":"1993","journal-title":"J Infect Dis"},{"key":"B20","doi-asserted-by":"publisher","first-page":"387","DOI":"10.1128\/CMR.00001-12","article-title":"The Cryptococcus neoformans capsule: a sword and a shield","volume":"25","author":"O\u2019Meara","year":"2012","journal-title":"Clin Microbiol Rev"},{"key":"B21","doi-asserted-by":"publisher","DOI":"10.1016\/j.mib.2023.102331","article-title":"Metals and the cell surface of Cryptococcus neoformans","volume":"74","author":"Xue","year":"2023","journal-title":"Curr Opin Microbiol"},{"key":"B22","doi-asserted-by":"publisher","DOI":"10.1016\/S0165-5728(02)00324-7","article-title":"Iron overload exacerbates experimental meningoencephalitis by Cryptococcus neoformans","volume":"132","author":"Barluzzi","year":"2002","journal-title":"J neuroimmunology"},{"key":"B23","doi-asserted-by":"publisher","first-page":"499","DOI":"10.1002\/lt.22024","article-title":"Pretransplant cryptococcosis and outcome after liver transplantation","volume":"16","author":"Sifri","year":"2010","journal-title":"Liver Transplant"},{"key":"B24","doi-asserted-by":"publisher","DOI":"10.1086\/656584","article-title":"Unrecognized pretransplant and donor-derived cryptococcal disease in organ transplant recipients","volume":"51","author":"Sun","year":"2010","journal-title":"Clin Infect Dis"},{"key":"B25","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0504823102","article-title":"HIV-1 Nef down-regulates the hemochromatosis protein HFE, manipulating cellular iron homeostasis","volume":"102","author":"Drakesmith","year":"2005","journal-title":"Proc Natl Acad Sci"},{"key":"B26","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1186\/s12967-017-1358-6","article-title":"The impact of antiretroviral therapy on iron homeostasis and inflammation markers in HIV-infected patients with mild anemia","volume":"15","author":"Quiros-Roldan","year":"2017","journal-title":"J Trans Med"},{"key":"B27","doi-asserted-by":"publisher","DOI":"10.1084\/jem.172.5.1293","article-title":"Prooxidant activity of transferrin and lactoferrin","volume":"172","author":"Klebanoff","year":"1990","journal-title":"J Exp Med"},{"key":"B28","doi-asserted-by":"publisher","first-page":"61","DOI":"10.1007\/s10534-008-9185-0","article-title":"Utilization of microbial iron assimilation processes for the development of new antibiotics and inspiration for the design of new anticancer agents","volume":"22","author":"Miller","year":"2009","journal-title":"Biometals"},{"key":"B29","doi-asserted-by":"publisher","DOI":"10.1517\/17460441.2012.708335","article-title":"Targeting iron assimilation to develop new antibacterials","volume":"7","author":"Foley","year":"2012","journal-title":"Expert Opin Drug Discov"},{"key":"B30","doi-asserted-by":"publisher","DOI":"10.2147\/dddt.s77860","article-title":"Effect of lactoferrin protein on red blood cells and macrophages: mechanism of parasite-host interaction","volume":"9","author":"Anand","year":"2015","journal-title":"Drug Des Devel Ther"},{"key":"B31","doi-asserted-by":"publisher","DOI":"10.1007\/s10534-009-9219-2","article-title":"Siderophores as drug delivery agents: application of the \u201cTrojan Horse\u201d strategy","volume":"22","author":"M\u00f6llmann","year":"2009","journal-title":"Biometals"},{"key":"B32","doi-asserted-by":"publisher","DOI":"10.1021\/ja109665t","article-title":"Design, synthesis, and study of a mycobactin\u2013 artemisinin conjugate that has selective and potent activity against tuberculosis and malaria","volume":"133","author":"Miller","year":"2011","journal-title":"J Am Chem Soc"},{"key":"B33","doi-asserted-by":"publisher","DOI":"10.1517\/13543784.10.2.309","article-title":"Antimicrobial properties of porphyrins","volume":"10","author":"Stojiljkovic","year":"2001","journal-title":"Expert Opin investigational Drugs"},{"key":"B34","doi-asserted-by":"publisher","DOI":"10.1093\/infdis\/jit029","article-title":"The mannoprotein Cig1 supports iron acquisition from heme and virulence in the pathogenic fungus Cryptococcus neoformans","volume":"207","author":"Cadieux","year":"2013","journal-title":"J Infect Dis"},{"key":"B35","doi-asserted-by":"publisher","DOI":"10.1016\/j.coph.2013.07.001","article-title":"Gallium-based anti-infectives: targeting microbial iron-uptake mechanisms","volume":"13","author":"Kelson","year":"2013","journal-title":"Curr Opin Pharmacol"},{"key":"B36","doi-asserted-by":"publisher","DOI":"10.1172\/JCI117119","article-title":"Chloroquine induces human macrophage killing of Histoplasma capsulatum by limiting the availability of intracellular iron and is therapeutic in a murine model of histoplasmosis","volume":"93","author":"Newman","year":"1994","journal-title":"J Clin Invest"},{"key":"B37","doi-asserted-by":"publisher","DOI":"10.1086\/315649","article-title":"Conditional lethality of the diprotic weak bases chloroquine and quinacrine against Cryptococcus neoformans","volume":"182","author":"Harrison","year":"2000","journal-title":"J Infect Dis"},{"key":"B38","doi-asserted-by":"publisher","DOI":"10.2147\/ijn.s85286","article-title":"Oral administration of encapsulated bovine lactoferrin protein nanocapsules against intracellular parasite Toxoplasma gondii","volume":"10","author":"Anand","year":"2015","journal-title":"Int J Nanomedicine"},{"key":"B39","doi-asserted-by":"publisher","first-page":"47","DOI":"10.2217\/nnm.15.181","article-title":"Antiparasitic and immunomodulatory potential of oral nanocapsules encapsulated lactoferrin protein against Plasmodium berghei","volume":"11","author":"Anand","year":"2016","journal-title":"Nanomedicine (Lond)"},{"key":"B40","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1000586","article-title":"Mucosal immunization with iron receptor antigens protects against urinary tract infection","volume":"5","author":"Alteri","year":"2009","journal-title":"PloS Pathog"},{"key":"B41","doi-asserted-by":"publisher","DOI":"10.1128\/aac.00892-10","article-title":"Fungicidal monoclonal antibody C7 interferes with iron acquisition in Candida albicans","volume":"55","author":"Brena","year":"2011","journal-title":"Antimicrob Agents Chemother"},{"key":"B42","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pbio.0040410","article-title":"Iron regulation of the major virulence factors in the AIDS-associated pathogen Cryptococcus neoformans","volume":"4","author":"Jung","year":"2006","journal-title":"PloS Biol"},{"key":"B43","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.0040045","article-title":"Iron source preference and regulation of iron uptake in Cryptococcus neoformans","volume":"4","author":"Jung","year":"2008","journal-title":"PloS Pathog"},{"key":"B44","doi-asserted-by":"publisher","DOI":"10.1128\/EC.00166-09","article-title":"Role of ferroxidases in iron uptake and virulence of Cryptococcus neoformans","volume":"8","author":"Jung","year":"2009","journal-title":"Eukaryotic Cell"},{"key":"B45","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.02377-18","article-title":"The monothiol glutaredoxin Grx4 regulates iron homeostasis and virulence in Cryptococcus neoformans","volume":"9","author":"Attarian","year":"2018","journal-title":"MBio"},{"key":"B46","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1001209","article-title":"HapX positively and negatively regulates the transcriptional response to iron deprivation in Cryptococcus neoformans","volume":"6","author":"Jung","year":"2010","journal-title":"PloS Pathog"},{"key":"B47","doi-asserted-by":"publisher","DOI":"10.1534\/genetics.120.303270","article-title":"A transcriptional regulatory map of iron homeostasis reveals a new control circuit for capsule formation in Cryptococcus neoformans","volume":"215","author":"Do","year":"2020","journal-title":"Genetics"},{"key":"B48","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1000776","article-title":"Interaction of Cryptococcus neoformans Rim101 and protein kinase A regulates capsule","volume":"6","author":"O\u2019Meara","year":"2010","journal-title":"PloS Pathog"},{"key":"B49","doi-asserted-by":"crossref","DOI":"10.1093\/infdis\/jit029","article-title":"The Mannoprotein Cig1 supports iron acquisition from heme and virulence in the pathogenic fungus Cryptococcus neoformans","volume-title":"J Infect Dis","author":"Cadieux","year":"2013"},{"key":"B50","doi-asserted-by":"publisher","DOI":"10.1016\/j.tim.2013.05.007","article-title":"An encapsulation of iron homeostasis and virulence in Cryptococcus neoformans","volume":"21","author":"Kronstad","year":"2013","journal-title":"Trends Microbiol"},{"key":"B51","doi-asserted-by":"publisher","DOI":"10.1128\/mbio.02944-22","article-title":"Unraveling the pathobiological role of the fungal KEOPS complex in Cryptococcus neoformans","volume":"13","author":"Choi","year":"2022","journal-title":"Mbio"},{"key":"B52","doi-asserted-by":"publisher","first-page":"1521","DOI":"10.1038\/s41467-020-15329-2","article-title":"Fungal kinases and transcription factors regulating brain infection in Cryptococcus neoformans","volume":"11","author":"Lee","year":"2020","journal-title":"Nat Commun"},{"key":"B53","doi-asserted-by":"publisher","DOI":"10.1016\/j.fgb.2004.08.001","article-title":"Double-joint PCR: a PCR-based molecular tool for gene manipulations in filamentous fungi","volume":"41","author":"Yu","year":"2004","journal-title":"Fungal Genet Biol"},{"key":"B54","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbrc.2009.10.089","article-title":"An efficient gene-disruption method in Cryptococcus neoformans by double-joint PCR with NAT-split markers","volume":"390","author":"Kim","year":"2009","journal-title":"Biochem Biophys Res Commun"},{"key":"B55","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-61779-539-8","article-title":"Host-fungus interactions: methods and protocols","author":"Brand","year":"2012","journal-title":"Host-Fungus Interactions."},{"key":"B56","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.01357-13","article-title":"Role of ferric reductases in iron acquisition and virulence in the fungal pathogen Cryptococcus neoformans","volume":"82","author":"Saikia","year":"2014","journal-title":"Infection Immun"},{"key":"B57","doi-asserted-by":"publisher","DOI":"10.1038\/75556","article-title":"Gene ontology: tool for the unification of biology. The Gene Ontology Consortium","volume":"25","author":"Ashburner","year":"2000","journal-title":"Nat Genet"},{"key":"B58","article-title":"GSEABase: gene set enrichment data structures and methods","volume-title":"R package version 1.56.0","author":"Martin","year":"2021"},{"key":"B59","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/btl567","article-title":"Using GOstats to test gene lists for GO term association","volume":"23","author":"Falcon","year":"2007","journal-title":"Bioinformatics"},{"key":"B60","article-title":"ggplot2; elegant graphics for data analysis, p 1\u2013213","volume-title":"Applied spatial data analysis R","author":"Wickham","year":"2009"},{"key":"B61","doi-asserted-by":"publisher","DOI":"10.1093\/mmy\/myx073","article-title":"The mitochondrial ABC transporter Atm1 plays a role in iron metabolism and virulence in the human fungal pathogen Cryptococcus neoformans","volume":"56","author":"Do","year":"2018","journal-title":"Med mycology"},{"key":"B62","doi-asserted-by":"publisher","first-page":"4339","DOI":"10.1038\/s41467-022-32006-8","article-title":"Structures of Atm1 provide insight into [2Fe-2S] cluster export from mitochondria","volume":"13","author":"Li","year":"2022","journal-title":"Nat Commun"},{"key":"B63","doi-asserted-by":"publisher","DOI":"10.1021\/bi7013272","article-title":"CGFS-type monothiol glutaredoxins from the cyanobacterium Synechocystis PCC6803 and other evolutionary distant model organisms possess a glutathione-ligated [2Fe-2S] cluster","volume":"46","author":"Picciocchi","year":"2007","journal-title":"Biochemistry"},{"key":"B64","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbrc.2011.04.069","article-title":"Multi-domain CGFS-type glutaredoxin Grx4 regulates iron homeostasis via direct interaction with a repressor Fep1 in fission yeast","volume":"408","author":"Kim","year":"2011","journal-title":"Biochem Biophys Res Commun"},{"key":"B65","doi-asserted-by":"publisher","first-page":"189","DOI":"10.1016\/j.cbpa.2020.02.008","article-title":"Iron\u2013sulfur cluster signaling: The common thread in fungal iron regulation","volume":"55","author":"Gupta","year":"2020","journal-title":"Curr Opin Chem Biol"},{"key":"B66","doi-asserted-by":"publisher","DOI":"10.1111\/j.1742-4658.2008.06721.x","article-title":"Phosphorylation of the Saccharomyces cerevisiae Grx4p glutaredoxin by the Bud32p kinase unveils a novel signaling pathway involving Sch9p, a yeast member of the Akt\/PKB subfamily","volume":"275","author":"Peggion","year":"2008","journal-title":"FEBS J"},{"key":"B67","doi-asserted-by":"publisher","first-page":"23","DOI":"10.1128\/EC.00181-12","article-title":"Evidence that two Pcl-like cyclins control Cdk9 activity during cell differentiation in Aspergillus nidulans asexual development","volume":"12","author":"Kempf","year":"2013","journal-title":"Eukaryotic Cell"},{"key":"B68","doi-asserted-by":"publisher","first-page":"e1002460","DOI":"10.1371\/journal.ppat.1002460","article-title":"Functional analysis of the kinome of the wheat scab fungus Fusarium graminearum","volume":"7","author":"Wang","year":"2011","journal-title":"PloS Pathog"}],"container-title":["Frontiers in Immunology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2025.1624237\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,7,25]],"date-time":"2025-07-25T11:19:39Z","timestamp":1753442379000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2025.1624237\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,7,25]]},"references-count":68,"alternative-id":["10.3389\/fimmu.2025.1624237"],"URL":"https:\/\/doi.org\/10.3389\/fimmu.2025.1624237","relation":{},"ISSN":["1664-3224"],"issn-type":[{"value":"1664-3224","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,7,25]]},"article-number":"1624237"}}