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As previously reported, the virulent behavior of this pathogen is enhanced by elevated levels of Ca\n                      <jats:sup>2+<\/jats:sup>\n                      that are commonly present in CF nasal and lung fluids. In addition, a Ca\n                      <jats:sup>2+<\/jats:sup>\n                      -binding EF-hand protein, EfhP (PA4107), was partially characterized and shown to be critical for the Ca\n                      <jats:sup>2+<\/jats:sup>\n                      -regulated virulence in\n                      <jats:italic>P. aeruginosa<\/jats:italic>\n                      . Here, we describe the rapid (10 min, 60 min), and adaptive (12 h) transcriptional responses of PAO1 to elevated Ca\n                      <jats:sup>2+<\/jats:sup>\n                      detected by genome-wide RNA sequencing and show that\n                      <jats:italic>efhP<\/jats:italic>\n                      deletion significantly hindered both rapid and adaptive Ca\n                      <jats:sup>2+<\/jats:sup>\n                      regulation. The most differentially regulated genes included multiple Fe sequestering mechanisms, a large number of extracytoplasmic function sigma factors (ECF\u03c3), and several virulence factors, such as the production of pyocins. The Ca\n                      <jats:sup>2+<\/jats:sup>\n                      regulation of Fe uptake was also observed in CF clinical isolates and appeared to involve the global regulator Fur. In addition, we showed that the\n                      <jats:italic>efhP<\/jats:italic>\n                      transcription is controlled by Ca\n                      <jats:sup>2+<\/jats:sup>\n                      and Fe, and this regulation required a Ca\n                      <jats:sup>2+<\/jats:sup>\n                      -dependent two-component regulatory system CarSR. Furthermore, the\n                      <jats:italic>efhP<\/jats:italic>\n                      expression is significantly increased in CF clinical isolates and upon pathogen internalization into epithelial cells. Overall, the results established for the first time that Ca\n                      <jats:sup>2+<\/jats:sup>\n                      controls Fe sequestering mechanisms in\n                      <jats:italic>P. aeruginosa<\/jats:italic>\n                      and that EfhP plays a key role in the regulatory interconnectedness between Ca\n                      <jats:sup>2+<\/jats:sup>\n                      and Fe signaling pathways, the two distinct and important signaling pathways that guide the pathogen\u2019s adaptation to the host.\n                    <\/jats:p>\n                    <jats:sec>\n                      <jats:title>IMPORTANCE<\/jats:title>\n                      <jats:p>\n                        <jats:italic>Pseudomonas aeruginosa<\/jats:italic>\n                        (\n                        <jats:italic>Pa<\/jats:italic>\n                        ) poses a major risk for severe infections, particularly in patients suffering from cystic fibrosis (CF). For the first time, kinetic RNA sequencing analysis identified\n                        <jats:italic>Pa<\/jats:italic>\n                        rapid and adaptive transcriptional responses to Ca\n                        <jats:sup>2+<\/jats:sup>\n                        levels consistent with those present in CF respiratory fluids. The most highly upregulated processes include iron sequestering, iron starvation sigma factors, and self-lysis factors pyocins. An EF-hand Ca\n                        <jats:sup>2+<\/jats:sup>\n                        sensor, EfhP, is required for at least 1\/3 of the Ca\n                        <jats:sup>2+<\/jats:sup>\n                        response, including the majority of the iron uptake mechanisms and the production of pyocins. Transcription of\n                        <jats:italic>efhP<\/jats:italic>\n                        itself is regulated by Ca\n                        <jats:sup>2+<\/jats:sup>\n                        and Fe, and increases during interactions with host epithelial cells, suggesting the protein\u2019s important role in\n                        <jats:italic>Pa<\/jats:italic>\n                        infections. The findings establish the regulatory interconnectedness between Ca\n                        <jats:sup>2+<\/jats:sup>\n                        and iron signaling pathways that shape\n                        <jats:italic>Pa<\/jats:italic>\n                        transcriptional responses. Therefore, understanding Pa\u2019s transcriptional response to Ca\n                        <jats:sup>2+<\/jats:sup>\n                        and associated regulatory mechanisms will serve in the development of future therapeutics targeting\n                        <jats:italic>Pa<\/jats:italic>\n                        \u2019s dangerous infections.\n                      <\/jats:p>\n                    <\/jats:sec>\n                  <\/jats:sec>","DOI":"10.1128\/mbio.02447-24","type":"journal-article","created":{"date-parts":[[2024,10,22]],"date-time":"2024-10-22T09:00:17Z","timestamp":1729587617000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":8,"title":["EF-hand calcium sensor, EfhP, controls transcriptional regulation of iron uptake by calcium in\n                    <i>Pseudomonas aeruginosa<\/i>"],"prefix":"10.1128","volume":"15","author":[{"given":"Jacob","family":"Burch-Konda","sequence":"first","affiliation":[{"name":"Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, Oklahoma, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Biraj B.","family":"Kayastha","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, Oklahoma, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Myriam","family":"Achour","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, Oklahoma, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aya","family":"Kubo","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, Oklahoma, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mackenzie","family":"Hull","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, Oklahoma, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Reygan","family":"Braga","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, Oklahoma, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lorelei","family":"Winton","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, Oklahoma, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rendi R.","family":"Rogers","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, Oklahoma, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Erika I.","family":"Lutter","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, Oklahoma, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0495-3867","authenticated-orcid":true,"given":"Marianna A.","family":"Patrauchan","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, Oklahoma, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_4_2_2","doi-asserted-by":"publisher","DOI":"10.1002\/ppul.10127"},{"key":"e_1_3_4_3_2","doi-asserted-by":"publisher","DOI":"10.1038\/srep10932"},{"key":"e_1_3_4_4_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmii.2014.08.019"},{"key":"e_1_3_4_5_2","volume-title":"Open forum infectious diseases","author":"Domitrovic TN","year":"2016","unstructured":"Domitrovic TN. 2016. 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