EF-hand protein, EfhP, specifically binds Ca2+ and mediates Ca2+ regulation of virulence in a human pathogen Pseudomonas aeruginosa
Abstract Calcium (Ca2+) is well known as a second messenger in eukaryotes, where Ca2+ signaling controls life-sustaining cellular processes. Although bacteria produce the components required for Ca2+ signaling, little is known about the mechanisms of bacterial Ca2+ signaling. Previously, we have ide...
| Main Authors: | , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2022-05-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-022-12584-9 |
| _version_ | 1828348157031874560 |
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| author | Biraj B. Kayastha Aya Kubo Jacob Burch-Konda Rosalie L. Dohmen Jacee L. McCoy Rendi R. Rogers Sergio Mares Justin Bevere Annalisa Huckaby William Witt Shuxia Peng Bharat Chaudhary Smita Mohanty Mariette Barbier Gabriel Cook Junpeng Deng Marianna A. Patrauchan |
| author_facet | Biraj B. Kayastha Aya Kubo Jacob Burch-Konda Rosalie L. Dohmen Jacee L. McCoy Rendi R. Rogers Sergio Mares Justin Bevere Annalisa Huckaby William Witt Shuxia Peng Bharat Chaudhary Smita Mohanty Mariette Barbier Gabriel Cook Junpeng Deng Marianna A. Patrauchan |
| author_sort | Biraj B. Kayastha |
| collection | DOAJ |
| description | Abstract Calcium (Ca2+) is well known as a second messenger in eukaryotes, where Ca2+ signaling controls life-sustaining cellular processes. Although bacteria produce the components required for Ca2+ signaling, little is known about the mechanisms of bacterial Ca2+ signaling. Previously, we have identified a putative Ca2+-binding protein EfhP (PA4107) with two canonical EF-hand motifs and reported that EfhP mediates Ca2+ regulation of virulence factors production and infectivity in Pseudomonas aeruginosa, a human pathogen causing life-threatening infections. Here, we show that EfhP selectively binds Ca2+ with 13.7 µM affinity, and that mutations at the +X and −Z positions within each or both EF-hand motifs abolished Ca2+ binding. We also show that the hydrophobicity of EfhP increased in a Ca2+ -dependent manner, however no such response was detected in the mutated proteins. 15 N-NMR showed Ca2+-dependent chemical shifts in EfhP confirming Ca2+-binding triggered structural rearrangements in the protein. Deletion of efhP impaired P. aeruginosa survival in macrophages and virulence in vivo. Disabling EfhP Ca2+ binding abolished Ca2+ induction of pyocyanin production in vitro. These data confirm that EfhP selectively binds Ca2+, which triggers its structural changes required for the Ca2+ regulation of P. aeruginosa virulence, thus establishing the role of EfhP as a Ca2+ sensor. |
| first_indexed | 2024-04-14T00:50:29Z |
| format | Article |
| id | doaj.art-25b2537cac2b4b44875de3dcda394a67 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-04-14T00:50:29Z |
| publishDate | 2022-05-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-25b2537cac2b4b44875de3dcda394a672022-12-22T02:21:49ZengNature PortfolioScientific Reports2045-23222022-05-0112111510.1038/s41598-022-12584-9EF-hand protein, EfhP, specifically binds Ca2+ and mediates Ca2+ regulation of virulence in a human pathogen Pseudomonas aeruginosaBiraj B. Kayastha0Aya Kubo1Jacob Burch-Konda2Rosalie L. Dohmen3Jacee L. McCoy4Rendi R. Rogers5Sergio Mares6Justin Bevere7Annalisa Huckaby8William Witt9Shuxia Peng10Bharat Chaudhary11Smita Mohanty12Mariette Barbier13Gabriel Cook14Junpeng Deng15Marianna A. Patrauchan16Department of Microbiology and Molecular Genetics, Oklahoma State UniversityDepartment of Microbiology and Molecular Genetics, Oklahoma State UniversityDepartment of Microbiology and Molecular Genetics, Oklahoma State UniversityDepartment of Microbiology and Molecular Genetics, Oklahoma State UniversityDepartment of Microbiology and Molecular Genetics, Oklahoma State UniversityDepartment of Microbiology and Molecular Genetics, Oklahoma State UniversityDepartment of Microbiology and Molecular Genetics, Oklahoma State UniversityVaccine Development Center at West Virginia UniversityVaccine Development Center at West Virginia UniversityVaccine Development Center at West Virginia UniversityDepartment of Biochemistry and Molecular Biology, Oklahoma State UniversityDepartment of Chemistry, Oklahoma State UniversityDepartment of Chemistry, Oklahoma State UniversityVaccine Development Center at West Virginia UniversityDepartment of Chemistry, Oklahoma State UniversityDepartment of Biochemistry and Molecular Biology, Oklahoma State UniversityDepartment of Microbiology and Molecular Genetics, Oklahoma State UniversityAbstract Calcium (Ca2+) is well known as a second messenger in eukaryotes, where Ca2+ signaling controls life-sustaining cellular processes. Although bacteria produce the components required for Ca2+ signaling, little is known about the mechanisms of bacterial Ca2+ signaling. Previously, we have identified a putative Ca2+-binding protein EfhP (PA4107) with two canonical EF-hand motifs and reported that EfhP mediates Ca2+ regulation of virulence factors production and infectivity in Pseudomonas aeruginosa, a human pathogen causing life-threatening infections. Here, we show that EfhP selectively binds Ca2+ with 13.7 µM affinity, and that mutations at the +X and −Z positions within each or both EF-hand motifs abolished Ca2+ binding. We also show that the hydrophobicity of EfhP increased in a Ca2+ -dependent manner, however no such response was detected in the mutated proteins. 15 N-NMR showed Ca2+-dependent chemical shifts in EfhP confirming Ca2+-binding triggered structural rearrangements in the protein. Deletion of efhP impaired P. aeruginosa survival in macrophages and virulence in vivo. Disabling EfhP Ca2+ binding abolished Ca2+ induction of pyocyanin production in vitro. These data confirm that EfhP selectively binds Ca2+, which triggers its structural changes required for the Ca2+ regulation of P. aeruginosa virulence, thus establishing the role of EfhP as a Ca2+ sensor.https://doi.org/10.1038/s41598-022-12584-9 |
| spellingShingle | Biraj B. Kayastha Aya Kubo Jacob Burch-Konda Rosalie L. Dohmen Jacee L. McCoy Rendi R. Rogers Sergio Mares Justin Bevere Annalisa Huckaby William Witt Shuxia Peng Bharat Chaudhary Smita Mohanty Mariette Barbier Gabriel Cook Junpeng Deng Marianna A. Patrauchan EF-hand protein, EfhP, specifically binds Ca2+ and mediates Ca2+ regulation of virulence in a human pathogen Pseudomonas aeruginosa Scientific Reports |
| title | EF-hand protein, EfhP, specifically binds Ca2+ and mediates Ca2+ regulation of virulence in a human pathogen Pseudomonas aeruginosa |
| title_full | EF-hand protein, EfhP, specifically binds Ca2+ and mediates Ca2+ regulation of virulence in a human pathogen Pseudomonas aeruginosa |
| title_fullStr | EF-hand protein, EfhP, specifically binds Ca2+ and mediates Ca2+ regulation of virulence in a human pathogen Pseudomonas aeruginosa |
| title_full_unstemmed | EF-hand protein, EfhP, specifically binds Ca2+ and mediates Ca2+ regulation of virulence in a human pathogen Pseudomonas aeruginosa |
| title_short | EF-hand protein, EfhP, specifically binds Ca2+ and mediates Ca2+ regulation of virulence in a human pathogen Pseudomonas aeruginosa |
| title_sort | ef hand protein efhp specifically binds ca2 and mediates ca2 regulation of virulence in a human pathogen pseudomonas aeruginosa |
| url | https://doi.org/10.1038/s41598-022-12584-9 |
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