{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T00:44:34Z","timestamp":1790037874588,"version":"4.0.1"},"reference-count":65,"publisher":"American Society for Microbiology","issue":"3","license":[{"start":{"date-parts":[[2009,3,1]],"date-time":"2009-03-01T00:00:00Z","timestamp":1235865600000},"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":["Eukaryot Cell"],"published-print":{"date-parts":[[2009,3]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            A library of more than 4,500 signature-tagged insertion mutants of the human pathogenic fungus\n            <jats:italic>Cryptococcus neoformans<\/jats:italic>\n            was generated, and a subset was screened in a murine inhalation model to identify genes required for virulence. New genes that regulate aspects of\n            <jats:italic>C. neoformans<\/jats:italic>\n            virulence were also identified by screening the entire library for in vitro phenotypes related to the ability to cause disease, including melanin production, growth at high temperature, and growth under conditions of nutrient limitation. A screen of 10% of the strain collection in mice identified an avirulent mutant strain with an insertion in the\n            <jats:italic>ENA1<\/jats:italic>\n            gene, which is predicted to encode a fungus-specific sodium or potassium P-type ATPase. The results of the deletion of the gene and complementation experiments confirmed its key role in mammalian virulence.\n            <jats:italic>ena1<\/jats:italic>\n            mutant strains exhibited no change in sensitivity to high salt concentrations but were sensitive to alkaline pH conditions, providing evidence that the fungus may have to survive at elevated pH during infection of the mammalian host. The mutation of the well-characterized virulence factor calcineurin (\n            <jats:italic>CNA1<\/jats:italic>\n            ) also rendered\n            <jats:italic>C. neoformans<\/jats:italic>\n            strains sensitive to elevated pH.\n            <jats:italic>ENA1<\/jats:italic>\n            transcripts in wild-type and\n            <jats:italic>cna1<\/jats:italic>\n            mutant strains were upregulated in response to high pH, and\n            <jats:italic>cna1 ena1<\/jats:italic>\n            double mutant strains exhibited increased sensitivity to elevated pH, indicating that at least two pathways in the fungus mediate survival under alkaline conditions. Signature-tagged mutagenesis is an effective strategy for the discovery of new virulence genes in fungal pathogens of animals.\n          <\/jats:p>","DOI":"10.1128\/ec.00375-08","type":"journal-article","created":{"date-parts":[[2009,1,17]],"date-time":"2009-01-17T03:01:56Z","timestamp":1232161316000},"page":"315-326","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":77,"title":["Identification of\n            <i>ENA1<\/i>\n            as a Virulence Gene of the Human Pathogenic Fungus\n            <i>Cryptococcus neoformans<\/i>\n            through Signature-Tagged Insertional Mutagenesis"],"prefix":"10.1128","volume":"8","author":[{"given":"Alexander","family":"Idnurm","sequence":"first","affiliation":[{"name":"Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, North Carolina 27710"},{"name":"Division of Cell Biology and Biophysics, School of Biological Sciences, University of Missouri\u2014Kansas City, Kansas City, Missouri 64110"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Felicia J.","family":"Walton","sequence":"additional","affiliation":[{"name":"Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, North Carolina 27710"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anna","family":"Floyd","sequence":"additional","affiliation":[{"name":"Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, North Carolina 27710"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jennifer L.","family":"Reedy","sequence":"additional","affiliation":[{"name":"Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, North Carolina 27710"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joseph","family":"Heitman","sequence":"additional","affiliation":[{"name":"Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, North Carolina 27710"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1091\/mbc.e04-11-0987"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.273.3.1640"},{"key":"e_1_3_2_4_2","first-page":"108","volume":"1","year":"2007","unstructured":"Bartlett, K. 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