{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,7]],"date-time":"2025-11-07T19:00:59Z","timestamp":1762542059431},"reference-count":39,"publisher":"American Society for Microbiology","issue":"8","license":[{"start":{"date-parts":[[2012,8,1]],"date-time":"2012-08-01T00:00:00Z","timestamp":1343779200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Antimicrob Agents Chemother"],"published-print":{"date-parts":[[2012,8]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            Systemic life-threatening fungal infections represent a significant unmet medical need. Cell-based, phenotypic screening can be an effective means of discovering potential novel antifungal compounds, but it does not address target identification, normally required for compound optimization by medicinal chemistry. Here, we demonstrate a combination of screening, genetic, and biochemical approaches to identify and characterize novel antifungal compounds. We isolated a set of novel non-azole antifungal compounds for which no target or mechanism of action is known, using a screen for inhibition of\n            <jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">Saccharomyces cerevisiae<\/jats:named-content>\n            proliferation. Haploinsufficiency profiling of these compounds in\n            <jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">S. cerevisiae<\/jats:named-content>\n            suggests that they target Erg11p, a cytochrome P450 family member, which is the target of azoles. Consistent with this, metabolic profiling in\n            <jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">S. cerevisiae<\/jats:named-content>\n            revealed a buildup of the metabolic intermediates prior to Erg11p activity, following compound treatment. Further, human cytochrome P450 is also inhibited in\n            <jats:italic>in vitro<\/jats:italic>\n            assays by these compounds. We modeled the Erg11p protein based on the human CYP51 crystal structure, and\n            <jats:italic>in silico<\/jats:italic>\n            docking of these compounds suggests that they interact with the heme center in a manner similar to that of azoles. Consistent with these docking observations,\n            <jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">Candida<\/jats:named-content>\n            strains carrying azole-resistant alleles of\n            <jats:italic>ERG11<\/jats:italic>\n            are also resistant to the compounds in this study. Thus, we have identified non-azole Erg11p inhibitors, using a systematic approach for ligand and target characterization.\n          <\/jats:p>","DOI":"10.1128\/aac.06332-11","type":"journal-article","created":{"date-parts":[[2012,5,22]],"date-time":"2012-05-22T02:55:06Z","timestamp":1337655306000},"page":"4233-4240","update-policy":"http:\/\/dx.doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":19,"title":["An Integrated Approach for Identification and Target Validation of Antifungal Compounds Active against Erg11p"],"prefix":"10.1128","volume":"56","author":[{"given":"Dominic","family":"Hoepfner","sequence":"first","affiliation":[{"name":"Novartis Institutes for BioMedical Research, Novartis Campus, Basel, Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shantanu","family":"Karkare","sequence":"additional","affiliation":[{"name":"Novartis Institutes for BioMedical Research, Novartis Campus, Basel, Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Stephen B.","family":"Helliwell","sequence":"additional","affiliation":[{"name":"Novartis Institutes for BioMedical Research, Novartis Campus, Basel, Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Martin","family":"Pfeifer","sequence":"additional","affiliation":[{"name":"Novartis Institutes for BioMedical Research, Novartis Campus, Basel, Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Markus","family":"Trunzer","sequence":"additional","affiliation":[{"name":"Novartis Institutes for BioMedical Research, Novartis Campus, Basel, Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sophie","family":"De Bonnechose","sequence":"additional","affiliation":[{"name":"Novartis Institutes for BioMedical Research, Novartis Campus, Basel, Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alfred","family":"Zimmerlin","sequence":"additional","affiliation":[{"name":"Novartis Institutes for BioMedical Research, Novartis Campus, Basel, Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jianshi","family":"Tao","sequence":"additional","affiliation":[{"name":"Novartis Institutes for BioMedical Research, Cambridge, Massachusetts, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Daryl","family":"Richie","sequence":"additional","affiliation":[{"name":"Novartis Institutes for BioMedical Research, Cambridge, Massachusetts, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Andreas","family":"Hofmann","sequence":"additional","affiliation":[{"name":"Novartis Institutes for BioMedical Research, Novartis Campus, Basel, Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Stefan","family":"Reinker","sequence":"additional","affiliation":[{"name":"Novartis Institutes for BioMedical Research, Novartis Campus, Basel, Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mathias","family":"Frederiksen","sequence":"additional","affiliation":[{"name":"Novartis Institutes for BioMedical Research, Novartis Campus, Basel, Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"N. Rao","family":"Movva","sequence":"additional","affiliation":[{"name":"Novartis Institutes for BioMedical Research, Novartis Campus, Basel, Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jeffrey A.","family":"Porter","sequence":"additional","affiliation":[{"name":"Novartis Institutes for BioMedical Research, Cambridge, Massachusetts, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Neil S.","family":"Ryder","sequence":"additional","affiliation":[{"name":"Novartis Institutes for BioMedical Research, Cambridge, Massachusetts, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Christian N.","family":"Parker","sequence":"additional","affiliation":[{"name":"Novartis Institutes for BioMedical Research, Novartis Campus, Basel, Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_3_2_2","doi-asserted-by":"crossref","first-page":"596","DOI":"10.1016\/0005-2760(81)90275-7","article-title":"Evidence for the presence of cytochrome P-450 functional in lanosterol 14 alpha-demethylation in microsomes of aerobically grown respiring yeast","volume":"665","author":"Aoyama Y","year":"1981","unstructured":"AoyamaY OkikawaT YoshidaY. 1981. Evidence for the presence of cytochrome P-450 functional in lanosterol 14 alpha-demethylation in microsomes of aerobically grown respiring yeast. Biochim. Biophys. Acta 665:596\u2013601.","journal-title":"Biochim. Biophys. Acta"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.46.4.943-946.2002"},{"key":"e_1_3_3_4_2","doi-asserted-by":"publisher","DOI":"10.1007\/BF02537115"},{"key":"e_1_3_3_5_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkp322"},{"key":"e_1_3_3_6_2","volume-title":"CLSI document M38-2A.","author":"CLSI","year":"2008","unstructured":"CLSI. 2008. Reference method for broth dilution antifungal susceptibility testing of filamentous fungi; approved standard, 2nd ed. CLSI document M38-2A. CLSI, Wayne, PA."},{"key":"e_1_3_3_7_2","volume-title":"CLSI document M27-A3.","author":"CLSI","year":"2008","unstructured":"CLSI. 2008. Reference method for broth dilution antifungal susceptibility testing of yeasts; approved standard, 3rd ed. CLSI document M27-A3. CLSI, Wayne PA."},{"key":"e_1_3_3_8_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.tim.2010.02.004"},{"key":"e_1_3_3_9_2","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1016\/S1359-6446(99)01430-0","article-title":"Emerging novel antifungal agents","volume":"5","author":"Fostel JM","year":"2000","unstructured":"FostelJM LarteyPA. 2000. Emerging novel antifungal agents. Drug Discov. Today 5:25\u201332.","journal-title":"Drug Discov. 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Biotechnol."},{"key":"e_1_3_3_16_2","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/16.6.566"},{"key":"e_1_3_3_17_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.tips.2008.07.006"},{"key":"e_1_3_3_18_2","doi-asserted-by":"crossref","first-page":"2675","DOI":"10.1093\/nar\/29.13.2675","article-title":"Comparison of the RNA polymerase III transcription machinery in Schizosaccharomyces pombe, Saccharomyces cerevisiae and human","volume":"29","author":"Huang Y","year":"2001","unstructured":"HuangY MaraiaRJ. 2001. Comparison of the RNA polymerase III transcription machinery in Schizosaccharomyces pombe, Saccharomyces cerevisiae and human. Nucleic Acids Res. 29:2675\u20132690.","journal-title":"Nucleic Acids Res."},{"key":"e_1_3_3_19_2","doi-asserted-by":"crossref","first-page":"351","DOI":"10.1002\/prot.10613","article-title":"A hierarchical approach to all-atom protein loop prediction","volume":"55","author":"Jacobson MP","year":"2004","unstructured":"JacobsonMP. 2004. A hierarchical approach to all-atom protein loop prediction. Proteins 55:351\u2013367.","journal-title":"Proteins"},{"key":"e_1_3_3_20_2","doi-asserted-by":"publisher","DOI":"10.1086\/367933"},{"key":"e_1_3_3_21_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.43.11.2798"},{"key":"e_1_3_3_22_2","doi-asserted-by":"crossref","first-page":"221","DOI":"10.1007\/BF02537824","article-title":"Cloning of the late genes in the ergosterol biosynthetic pathway of Saccharomyces cerevisiae\u2014a review","volume":"30","author":"Lees N","year":"1995","unstructured":"LeesN SkaggsB KirschD BardM. 1995. Cloning of the late genes in the ergosterol biosynthetic pathway of Saccharomyces cerevisiae\u2014a review. Lipids 30:221\u2013226.","journal-title":"Lipids"},{"key":"e_1_3_3_23_2","first-page":"33","article-title":"Biochemistry and molecular biology of sterol synthesis in Saccharomyces cerevisiae","volume":"34","author":"Lees NDD","year":"1999","unstructured":"LeesNDD. 1999. 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Miconazole induces changes in actin cytoskeleton prior to reactive oxygen species induction in yeast. J. Biol. Chem. 282:21592\u201321597.","journal-title":"J. Biol. 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