{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,31]],"date-time":"2026-01-31T00:58:44Z","timestamp":1769821124843,"version":"3.49.0"},"reference-count":50,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2023,2,10]],"date-time":"2023-02-10T00:00:00Z","timestamp":1675987200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"JSPS KAKENHI","award":["20K07475"],"award-info":[{"award-number":["20K07475"]}]},{"name":"JSPS KAKENHI","award":["18H02653"],"award-info":[{"award-number":["18H02653"]}]},{"name":"Hamaguchi Foundation","award":["20K07475"],"award-info":[{"award-number":["20K07475"]}]},{"name":"Hamaguchi Foundation","award":["18H02653"],"award-info":[{"award-number":["18H02653"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["JoF"],"abstract":"<jats:p>Mitochondrial dysfunction or morphological abnormalities in human pathogenic fungi are known to contribute to azole resistance; however, the underlying molecular mechanisms are unknown. In this study, we investigated the link between mitochondrial morphology and azole resistance in Candida glabrata, which is the second most common cause of human candidiasis worldwide. The ER-mitochondrial encounter structure (ERMES) complex is thought to play an important role in the mitochondrial dynamics necessary for mitochondria to maintain their function. Of the five components of the ERMES complex, deletion of GEM1 increased azole resistance. Gem1 is a GTPase that regulates the ERMES complex activity. Point mutations in GEM1 GTPase domains were sufficient to confer azole resistance. The cells lacking GEM1 displayed abnormalities in mitochondrial morphology, increased mtROS levels, and increased expression of azole drug efflux pumps encoded by CDR1 and CDR2. Interestingly, treatment with N-acetylcysteine (NAC), an antioxidant, reduced ROS production and the expression of CDR1 in \u0394gem1 cells. Altogether, the absence of Gem1 activity caused an increase in mitochondrial ROS concentration, leading to Pdr1-dependent upregulation of the drug efflux pump Cdr1, resulting in azole resistance.<\/jats:p>","DOI":"10.3390\/jof9020240","type":"journal-article","created":{"date-parts":[[2023,2,13]],"date-time":"2023-02-13T02:42:33Z","timestamp":1676256153000},"page":"240","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["In Candida glabrata, ERMES Component GEM1 Controls Mitochondrial Morphology, mtROS, and Drug Efflux Pump Expression, Resulting in Azole Susceptibility"],"prefix":"10.3390","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4280-6048","authenticated-orcid":false,"given":"Michiyo","family":"Okamoto","sequence":"first","affiliation":[{"name":"Medical Mycology Research Center, Chiba University, Chiba 260-8673, Japan"}]},{"given":"Keiko","family":"Nakano","sequence":"additional","affiliation":[{"name":"Medical Mycology Research Center, Chiba University, Chiba 260-8673, Japan"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6652-4654","authenticated-orcid":false,"given":"Azusa","family":"Takahashi-Nakaguchi","sequence":"additional","affiliation":[{"name":"Medical Mycology Research Center, Chiba University, Chiba 260-8673, Japan"}]},{"given":"Kaname","family":"Sasamoto","sequence":"additional","affiliation":[{"name":"Medical Mycology Research Center, Chiba University, Chiba 260-8673, Japan"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1903-1555","authenticated-orcid":false,"given":"Masashi","family":"Yamaguchi","sequence":"additional","affiliation":[{"name":"Medical Mycology Research Center, Chiba University, Chiba 260-8673, Japan"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5676-6174","authenticated-orcid":false,"given":"Miguel Cacho","family":"Teixeira","sequence":"additional","affiliation":[{"name":"iBB\u2014Institute for Bioengineering and Biosciences, Instituto Superior T\u00e9cnico, Universidade de Lisboa, 1049-001 Lisbon, Portugal"},{"name":"Associate Laboratory i4HB\u2014Institute for Health and Bioeconomy at Instituto Superior T\u00e9cnico, Universidade de Lisboa, 1049-001 Lisbon, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3400-5569","authenticated-orcid":false,"given":"Hiroji","family":"Chibana","sequence":"additional","affiliation":[{"name":"Medical Mycology Research Center, Chiba University, Chiba 260-8673, Japan"}]}],"member":"1968","published-online":{"date-parts":[[2023,2,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"133","DOI":"10.1128\/CMR.00029-06","article-title":"Epidemiology of Invasive Candidiasis: A Persistent Public Health Problem","volume":"20","author":"Pfaller","year":"2007","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1445","DOI":"10.1056\/NEJMra1315399","article-title":"Invasive Candidiasis","volume":"373","author":"Kullberg","year":"2015","journal-title":"N. Engl. J. Med."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1111\/1469-0691.12539","article-title":"Global Trends in the Distribution of Candida Species Causing Candidemia","volume":"20","author":"Guinea","year":"2014","journal-title":"Clin. Microbiol. Infect."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1016\/j.diagmicrobio.2012.02.001","article-title":"The Changing Epidemiology of Healthcare-Associated Candidemia over Three Decades","volume":"73","author":"Diekema","year":"2012","journal-title":"Diagn Microbiol. Infect Dis."},{"key":"ref_5","first-page":"e449","article-title":"Burden of Candidemia in the United States, 2017","volume":"71","author":"Tsay","year":"2020","journal-title":"Clin. Infect. Dis."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"e01564-17","DOI":"10.1128\/JCM.01564-17","article-title":"Update from a 12-Year Nationwide Fungemia Surveillance: Increasing Intrinsic and Acquired Resistance Causes Concern","volume":"56","author":"Astvad","year":"2018","journal-title":"J. Clin. Microbiol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1103","DOI":"10.1093\/jac\/dkx047","article-title":"Changing Epidemiology of Candidaemia in Australia","volume":"72","author":"Chapman","year":"2017","journal-title":"J. Antimicrob. Chemother."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"701","DOI":"10.1111\/cmi.12091","article-title":"Two Unlike Cousins: Candida albicans and C. glabrata Infection Strategies","volume":"15","author":"Brunke","year":"2013","journal-title":"Cell Microbiol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"910","DOI":"10.1006\/bbrc.1995.1272","article-title":"Mode of Action and Resistance to Azole Antifungals Associated with the Formation of 14\u03b1-Methylergosta-8,24(28)-Dien-3\u03b2,6\u03b1-Diol","volume":"207","author":"Kelly","year":"1995","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"2753","DOI":"10.1128\/AAC.43.11.2753","article-title":"The ATP Binding Cassette Transporter Gene CgCDR1 from Candida glabrata Is Involved in the Resistance of Clinical Isolates to Azole Antifungal Agents","volume":"43","author":"Sanglard","year":"1999","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1174","DOI":"10.1128\/AAC.45.4.1174-1183.2001","article-title":"Role of ATP-Binding-Cassette Transporter Genes in High-Frequency Acquisition of Resistance to Azole Antifungals in Candida glabrata","volume":"45","author":"Sanglard","year":"2001","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"3773","DOI":"10.1128\/AAC.48.10.3773-3781.2004","article-title":"Azole Resistance in Candida glabrata: Coordinate Upregulation of Multidrug Transporters and Evidence for a Pdr1-Like Transcription Factor","volume":"48","author":"Vermitsky","year":"2004","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1384","DOI":"10.1128\/AAC.50.4.1384-1392.2006","article-title":"Candida glabrata PDR1, a Transcriptional Regulator of a Pleiotropic Drug Resistance Network, Mediates Azole Resistance in Clinical Isolates and Petite Mutants","volume":"50","author":"Tsai","year":"2006","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1852","DOI":"10.1128\/AAC.01271-10","article-title":"Loss of Mitochondrial Functions Associated with Azole Resistance in Candida glabrata Results in Enhanced Virulence in Mice","volume":"55","author":"Ferrari","year":"2011","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"977","DOI":"10.1099\/0022-1317-49-11-977","article-title":"In-Vivo Selection of an Azole-Resistant Petite Mutant of Candida glabrata","volume":"49","author":"Bouchara","year":"2000","journal-title":"J. Med. Microbiol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"3046","DOI":"10.1128\/JCM.00526-06","article-title":"Azole Resistance of Candida glabrata in a Case of Recurrent Fungemia","volume":"44","author":"Posteraro","year":"2006","journal-title":"J. Clin. Microbiol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1600","DOI":"10.1128\/AAC.48.5.1600-1613.2004","article-title":"Functional Genomic Analysis of Fluconazole Susceptibility in the Pathogenic Yeast Candida glabrata: Roles of Calcium Signaling and Mitochondria","volume":"48","author":"Kaur","year":"2004","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"930","DOI":"10.1111\/mmi.13167","article-title":"Mitochondrial Dynamics in the Pathogenic Mold Aspergillus fumigatus: Therapeutic and Evolutionary Implications","volume":"98","author":"Neubauer","year":"2015","journal-title":"Mol. Microbiol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"616","DOI":"10.1016\/j.tibs.2011.08.004","article-title":"Staying in Touch: The Molecular Era of Organelle Contact Sites","volume":"36","author":"Elbaz","year":"2011","journal-title":"Trends Biochem. Sci."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"2526","DOI":"10.1016\/j.bbamcr.2013.01.028","article-title":"Organization and Function of Membrane Contact Sites","volume":"1833","author":"Helle","year":"2013","journal-title":"Biochim. Et Biophys. Acta BBA-Mol. Cell Res."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"477","DOI":"10.1126\/science.1175088","article-title":"An ER-Mitochondria Tethering Complex Revealed by a Synthetic Biology Screen","volume":"325","author":"Kornmann","year":"2009","journal-title":"Science"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"14151","DOI":"10.1073\/pnas.1111314108","article-title":"The Conserved GTPase Gem1 Regulates Endoplasmic Reticulum\u2013Mitochondria Connections","volume":"108","author":"Kornmann","year":"2011","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"743","DOI":"10.1016\/j.jmb.2011.09.012","article-title":"Composition and Topology of the Endoplasmic Reticulum\u2013Mitochondria Encounter Structure","volume":"413","author":"Stroud","year":"2011","journal-title":"J. Mol. Biol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1927","DOI":"10.1093\/bioinformatics\/btq326","article-title":"Homology of SMP Domains to the TULIP Superfamily of Lipid-Binding Proteins Provides a Structural Basis for Lipid Exchange between ER and Mitochondria","volume":"26","author":"Kopec","year":"2010","journal-title":"Bioinformatics"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"959","DOI":"10.1083\/jcb.201704119","article-title":"Structure\u2013Function Insights into Direct Lipid Transfer between Membranes by Mmm1\u2013Mdm12 of ERMES","volume":"217","author":"Kawano","year":"2018","journal-title":"J. Cell Biol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"30777","DOI":"10.1038\/srep30777","article-title":"A Phospholipid Transfer Function of ER-Mitochondria Encounter Structure Revealed in Vitro","volume":"6","author":"Kojima","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"545","DOI":"10.1083\/jcb.136.3.545","article-title":"Mdm12p, a Component Required for Mitochondrial Inheritance That Is Conserved between Budding and Fission Yeast","volume":"136","author":"Berger","year":"1997","journal-title":"J. Cell Biol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1375","DOI":"10.1083\/jcb.126.6.1375","article-title":"MMM1 Encodes a Mitochondrial Outer Membrane Protein Essential for Establishing and Maintaining the Structure of Yeast Mitochondria","volume":"126","author":"Burgess","year":"1994","journal-title":"J. Cell Biol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1361","DOI":"10.1083\/jcb.126.6.1361","article-title":"Regulation of Mitochondrial Morphology and Inheritance by Mdm10p, a Protein of the Mitochondrial Outer Membrane","volume":"126","author":"Sogo","year":"1994","journal-title":"J. Cell Biol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"677","DOI":"10.1083\/jcb.200308012","article-title":"Mmm2p, a Mitochondrial Outer Membrane Protein Required for Yeast Mitochondrial Shape and Maintenance of mtDNA Nucleoids","volume":"164","author":"Youngman","year":"2004","journal-title":"J. Cell Biol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"4618","DOI":"10.1091\/mbc.e03-04-0225","article-title":"A Protein Complex Containing Mdm10p, Mdm12p, and Mmm1p Links Mitochondrial Membranes and DNA to the Cytoskeleton-Based Segregation Machinery","volume":"14","author":"Boldogh","year":"2003","journal-title":"Mol. Biol. Cell"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"450","DOI":"10.1016\/j.devcel.2014.01.012","article-title":"Mitochondrial ER Contacts Are Crucial for Mitophagy in Yeast","volume":"28","author":"Westermann","year":"2014","journal-title":"Dev. Cell"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"e00422","DOI":"10.7554\/eLife.00422","article-title":"ER-Associated Mitochondrial Division Links the Distribution of Mitochondria and Mitochondrial DNA in Yeast","volume":"2","author":"Murley","year":"2013","journal-title":"eLife"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"2555","DOI":"10.3389\/fmicb.2017.02555","article-title":"The Mitochondrial GTPase Gem1 Contributes to the Cell Wall Stress Response and Invasive Growth of Candida albicans","volume":"8","author":"Koch","year":"2017","journal-title":"Front. Microbiol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"e00074-16","DOI":"10.1128\/mSphere.00074-16","article-title":"The Endoplasmic Reticulum-Mitochondrion Tether ERMES Orchestrates Fungal Immune Evasion, Illuminating Inflammasome Responses to Hyphal Signals","volume":"1","author":"Tucey","year":"2016","journal-title":"Msphere"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1016\/j.ijmm.2016.11.005","article-title":"The ER-Mitochondria Encounter Structure Contributes to Hyphal Growth, Mitochondrial Morphology and Virulence of the Pathogenic Mold Aspergillus fumigatus","volume":"307","author":"Penka","year":"2017","journal-title":"Int. J. Med. Microbiol."},{"key":"ref_37","first-page":"67","article-title":"Requirement of a Putative Mitochondrial GTPase, GemA, for Azole Susceptibility, Virulence, and Cell Wall Integrity in Aspergillus fumigatus","volume":"13","author":"Zhou","year":"2022","journal-title":"Front. Microbiol."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Ueno, K., Matsumoto, Y., Uno, J., Sasamoto, K., Sekimizu, K., Kinjo, Y., and Chibana, H. (2011). Intestinal Resident Yeast Candida glabrata Requires Cyb2p-Mediated Lactate Assimilation to Adapt in Mouse Intestine. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0024759"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"e00099-18","DOI":"10.1128\/mSphere.00099-18","article-title":"Construction and Use of a Recyclable Marker to Examine the Role of Major Facilitator Superfamily Protein Members in Candida glabrata Drug Resistance Phenotypes","volume":"3","author":"Vu","year":"2018","journal-title":"Msphere"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"820675","DOI":"10.3389\/fcell.2022.820675","article-title":"Erg25 Controls Host-Cholesterol Uptake Mediated by Aus1p-Associated Sterol-Rich Membrane Domains in Candida glabrata","volume":"10","author":"Okamoto","year":"2022","journal-title":"Front. Cell Dev. Biol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1675","DOI":"10.1534\/g3.113.006908","article-title":"Expression Plasmids for Use in Candida glabrata","volume":"3","author":"Zordan","year":"2013","journal-title":"G3 Genes Genomes Genet."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"354","DOI":"10.1074\/jbc.M110.180034","article-title":"Structure-Function Analysis of the Yeast Mitochondrial Rho GTPase, Gem1p: Implications for Mitochondrial Inheritance","volume":"286","author":"Koshiba","year":"2011","journal-title":"J. Biol. Chem."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1769","DOI":"10.1021\/acschembio.7b00339","article-title":"Real-Time Imaging of the Azole Class of Antifungal Drugs in Live Candida Cells","volume":"12","author":"Benhamou","year":"2017","journal-title":"ACS Chem. Biol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"358","DOI":"10.1126\/science.1207385","article-title":"ER Tubules Mark Sites of Mitochondrial Division","volume":"334","author":"Friedman","year":"2011","journal-title":"Science"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"1147","DOI":"10.1093\/genetics\/162.3.1147","article-title":"Maintenance of Mitochondrial Morphology Is Linked to Maintenance of the Mitochondrial Genome in Saccharomyces cerevisiae","volume":"162","author":"Hanekamp","year":"2002","journal-title":"Genetics"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"e00934-19","DOI":"10.1128\/mBio.00934-19","article-title":"Evidence That Ergosterol Biosynthesis Modulates Activity of the Pdr1 Transcription Factor in Candida glabrata","volume":"10","author":"Vu","year":"2019","journal-title":"MBio"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"641","DOI":"10.1007\/s12223-022-00968-3","article-title":"The UPC2 Gene in Kluyveromyces lactis Stress Adaptation","volume":"67","author":"Betinova","year":"2022","journal-title":"Folia Microbiol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"11863","DOI":"10.1073\/pnas.1301951110","article-title":"Dynamics of Nucleoid Structure Regulated by Mitochondrial Fission Contributes to Cristae Reformation and Release of Cytochrome c","volume":"110","author":"Ishihara","year":"2013","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"203","DOI":"10.1016\/0378-1119(95)00552-H","article-title":"Cloning of the Candida glabrata TRP1 and HIS3 Genes, and Construction of Their Disruptant Strains by Sequential Integrative Transformation","volume":"165","author":"Kitada","year":"1995","journal-title":"Gene"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"1239","DOI":"10.1128\/EC.00414-06","article-title":"Development of a Highly Efficient Gene Targeting System Induced by Transient Repression of YKU80 Expression in Candida glabrata","volume":"6","author":"Ueno","year":"2007","journal-title":"Eukaryotic Cell"}],"container-title":["Journal of Fungi"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2309-608X\/9\/2\/240\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T18:31:08Z","timestamp":1760121068000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2309-608X\/9\/2\/240"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,2,10]]},"references-count":50,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2023,2]]}},"alternative-id":["jof9020240"],"URL":"https:\/\/doi.org\/10.3390\/jof9020240","relation":{},"ISSN":["2309-608X"],"issn-type":[{"value":"2309-608X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,2,10]]}}}