{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,7,30]],"date-time":"2025-07-30T12:59:56Z","timestamp":1753880396346,"version":"3.41.2"},"reference-count":42,"publisher":"American Society for Microbiology","issue":"1","license":[{"start":{"date-parts":[[2017,2,7]],"date-time":"2017-02-07T00:00:00Z","timestamp":1486425600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2017,3,8]],"date-time":"2017-03-08T00:00:00Z","timestamp":1488931200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/100000060","name":"HHS | NIH | National Institute of Allergy and Infectious Diseases","doi-asserted-by":"publisher","award":["R01 AI112595"],"award-info":[{"award-number":["R01 AI112595"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000060","name":"HHS | NIH | National Institute of Allergy and Infectious Diseases","doi-asserted-by":"publisher","award":["T32 AI052080"],"award-info":[{"award-number":["T32 AI052080"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["mBio"],"published-print":{"date-parts":[[2017,3,8]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            Invasive aspergillosis (IA), caused by the filamentous fungal pathogen\n            <jats:italic>Aspergillus fumigatus<\/jats:italic>\n            , is a major cause of death among immunocompromised patients. The cyclic AMP\/protein kinase A (PKA) signaling pathway is essential for hyphal growth and virulence of\n            <jats:italic>A.\u00a0fumigatus<\/jats:italic>\n            , but the mechanism of regulation of PKA remains largely unknown. Here, we discovered a novel mechanism for the regulation of PKA activity in\n            <jats:italic>A.\u00a0fumigatus<\/jats:italic>\n            via phosphorylation of key residues within the major catalytic subunit, PkaC1. Phosphopeptide enrichment and tandem mass spectrometry revealed the phosphorylation of PkaC1 at four sites (S175, T331, T333, and T337) with implications for important and diverse roles in the regulation of\n            <jats:italic>A.\u00a0fumigatus<\/jats:italic>\n            PKA. While the phosphorylation at one of the residues (T333) is conserved in other species, the identification of three other residues represents previously unknown PKA phosphoregulation in\n            <jats:italic>A.\u00a0fumigatus<\/jats:italic>\n            . Site-directed mutagenesis of the phosphorylated residues to mimic or prevent phosphorylation revealed dramatic effects on kinase activity, growth, conidiation, cell wall stress response, and virulence in both invertebrate and murine infection models. Three-dimensional structural modeling of\n            <jats:italic>A.\u00a0fumigatus<\/jats:italic>\n            PkaC1 substantiated the positive or negative regulatory roles for specific residues. Suppression of PKA activity also led to downregulation of PkaC1 protein levels in an apparent novel negative-feedback mechanism. Taken together, we propose a model in which PkaC1 phosphorylation both positively and negatively modulates its activity. These findings pave the way for future discovery of fungus-specific aspects of this key signaling network.\n          <\/jats:p>\n          <jats:p>\n            <jats:bold>IMPORTANCE<\/jats:bold>\n            Our understanding of signal transduction networks in pathogenic fungi is limited, despite the increase in invasive fungal infections and rising mortality rates in the immunosuppressed patient population. Because PKA is known to be essential for hyphal growth and virulence of\n            <jats:italic>A.\u00a0fumigatus<\/jats:italic>\n            , we sought to identify fungus-specific regulatory mechanisms governing PKA activity. In this study, we identify, for the first time, a novel mechanism for the regulation of PKA signaling in which differential phosphorylation of the PkaC1 catalytic subunit can lead to either positive or negative regulation of activity. Furthermore, we show that inactivation of PKA signaling leads to downregulation of catalytic subunit protein levels in a negative-feedback mechanism distinct from expression patterns previously reported in the yeasts. Our findings represent a divergence in the regulation of PKA signaling in\n            <jats:italic>A.\u00a0fumigatus<\/jats:italic>\n            , which could potentially be exploited as a target and also open the avenue for discovery of fungus-specific downstream effectors of PKA.\n          <\/jats:p>","DOI":"10.1128\/mbio.02319-16","type":"journal-article","created":{"date-parts":[[2017,2,8]],"date-time":"2017-02-08T01:10:32Z","timestamp":1486516232000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":9,"title":["A Novel Phosphoregulatory Switch Controls the Activity and Function of the Major Catalytic Subunit of Protein Kinase A in\n            <i>Aspergillus fumigatus<\/i>"],"prefix":"10.1128","volume":"8","author":[{"given":"E. Keats","family":"Shwab","sequence":"first","affiliation":[{"name":"Division of Pediatric Infectious Diseases, Department of Pediatrics, Duke University Medical Center, Durham, North Carolina, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Praveen R.","family":"Juvvadi","sequence":"additional","affiliation":[{"name":"Division of Pediatric Infectious Diseases, Department of Pediatrics, Duke University Medical Center, Durham, North Carolina, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Greg","family":"Waitt","sequence":"additional","affiliation":[{"name":"Duke Proteomics and Metabolomics Core Facility, Center for Genomic and Computational Biology, Duke University, Durham, North Carolina, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Erik J.","family":"Soderblom","sequence":"additional","affiliation":[{"name":"Duke Proteomics and Metabolomics Core Facility, Center for Genomic and Computational Biology, Duke University, Durham, North Carolina, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"M. Arthur","family":"Moseley","sequence":"additional","affiliation":[{"name":"Duke Proteomics and Metabolomics Core Facility, Center for Genomic and Computational Biology, Duke University, Durham, North Carolina, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nathan I.","family":"Nicely","sequence":"additional","affiliation":[{"name":"Duke Human Vaccine Institute, Duke University School of Medicine, Durham, North Carolina, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yohannes G.","family":"Asfaw","sequence":"additional","affiliation":[{"name":"Department of Laboratory Animal Resources, Duke University Medical Center, Durham, North Carolina, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"William J.","family":"Steinbach","sequence":"additional","affiliation":[{"name":"Division of Pediatric Infectious Diseases, Department of Pediatrics, Duke University Medical Center, Durham, North Carolina, USA"},{"name":"Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, North Carolina, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.2741\/A195"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.1007\/s002030050659"},{"key":"e_1_3_3_4_2","doi-asserted-by":"publisher","DOI":"10.1080\/10408410802645646"},{"key":"e_1_3_3_5_2","first-page":"1068","article-title":"The epidemiology of fungal infections in patients with hematologic malignancies: the SEIFEM-2004 study","volume":"91","author":"Pagano L","year":"2006","unstructured":"Pagano L Caira M Candoni A Offidani M Fianchi L Martino B Pastore D Picardi M Bonini A Chierichini A Fanci R Caramatti C Invernizzi R Mattei D Mitra ME Melillo L Aversa F Van Lint MT Falcucci P Valentini CG Girmenia C Nosari A . 2006. The epidemiology of fungal infections in patients with hematologic malignancies: the SEIFEM-2004 study. 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