{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,4,7]],"date-time":"2026-04-07T22:37:48Z","timestamp":1775601468113,"version":"3.50.1"},"posted":{"date-parts":[[2024,1,10]]},"group-title":"Microbiology","reference-count":118,"publisher":"openRxiv","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2024,1,10]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:p>\n                  The human pathogen\n                  <jats:italic>Pseudomonas aeruginosa<\/jats:italic>\n                  poses a major risk for a range of severe infections, particularly lung infections in patients suffering from cystic fibrosis (CF). 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 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 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 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 all the iron uptake mechanisms and production of pyocins. Transcription of\n                    <jats:italic>efhP<\/jats:italic>\n                    itself is regulated by Ca\n                    <jats:sup>2+<\/jats:sup>\n                    , 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 the development of future therapeutics targeting\n                    <jats:italic>Pa<\/jats:italic>\n                    dangerous infections.\n                  <\/jats:p>\n                <\/jats:sec>","DOI":"10.1101\/2024.01.09.574892","type":"posted-content","created":{"date-parts":[[2024,1,10]],"date-time":"2024-01-10T13:15:29Z","timestamp":1704892529000},"source":"Crossref","is-referenced-by-count":1,"title":["EF-Hand Calcium Sensor, EfhP, Controls Transcriptional Regulation of Iron Uptake by Calcium in\n                  <i>Pseudomonas aeruginosa<\/i>"],"prefix":"10.64898","author":[{"given":"Jacob","family":"Burch-Konda","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Biraj B.","family":"Kayastha","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Aya","family":"Kubo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Myriam","family":"Achour","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mackenzie","family":"Hull","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Reygan","family":"Braga","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lorelei","family":"Winton","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rendi R.","family":"Rogers","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jacee","family":"McCoy","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Erika I.","family":"Lutter","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0495-3867","authenticated-orcid":false,"given":"Marianna A.","family":"Patrauchan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2024011121400594000_2024.01.09.574892v1.1","doi-asserted-by":"publisher","DOI":"10.1002\/ppul.10127"},{"key":"2024011121400594000_2024.01.09.574892v1.2","doi-asserted-by":"crossref","first-page":"10932","DOI":"10.1038\/srep10932","article-title":"Phenotypic diversity within a Pseudomonas aeruginosa population infecting an adult with cystic fibrosis","volume":"5","year":"2015","journal-title":"Scientific reports"},{"key":"2024011121400594000_2024.01.09.574892v1.3","first-page":"516","article-title":"Pseudomonas aeruginosa infective endocarditis in patients who do not use intravenous drugs: analysis of risk factors and treatment outcomes. 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