{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T13:31:20Z","timestamp":1768483880296,"version":"3.49.0"},"reference-count":42,"publisher":"Oxford University Press (OUP)","issue":"8","license":[{"start":{"date-parts":[[2023,6,8]],"date-time":"2023-06-08T00:00:00Z","timestamp":1686182400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/pages\/standard-publication-reuse-rights"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["21977037"],"award-info":[{"award-number":["21977037"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003453","name":"Guangdong Natural Science Foundation","doi-asserted-by":"publisher","award":["2022A1515010674"],"award-info":[{"award-number":["2022A1515010674"]}],"id":[{"id":"10.13039\/501100003453","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100012166","name":"National Key R & D Program of China","doi-asserted-by":"crossref","award":["2017YFA0505100"],"award-info":[{"award-number":["2017YFA0505100"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100004493","name":"International Atomic Energy Agency","doi-asserted-by":"publisher","award":["25076"],"award-info":[{"award-number":["25076"]}],"id":[{"id":"10.13039\/501100004493","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2023,8,2]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:sec>\n                  <jats:title>Background<\/jats:title>\n                  <jats:p>CpxR is a critical regulator in bacterial adaptation to various harmful stresses, and is known to regulate bacterial resistance to commonly used antibiotics, such as aminoglycosides, \u03b2-lactams and polypeptides. However, the detailed study of functional residues of CpxR remains insufficient.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Objectives<\/jats:title>\n                  <jats:p>To investigate the contribution of Lys219 to CpxR\u2019s function in regulating antibiotic resistance of Escherichia coli.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Methods<\/jats:title>\n                  <jats:p>We performed sequence alignment and conservative analysis of the CpxR protein and constructed mutant strains. We then performed electrophoretic mobility shift assay, real-time quantitative PCR assay, determination of reactive oxygen species (ROS) levels, molecular dynamics simulation, conformational analysis and circular dichroism.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Results<\/jats:title>\n                  <jats:p>All mutant proteins (K219Q, K219A and K219R) lost the cpxP DNA-binding ability. Additionally, the three complemented strains eK219A, eK219Q, and eK219R exhibited lower resistance to copper toxicity and alkaline pH toxicity than eWT. Molecular dynamics analysis revealed that mutation of Lys219 leads to looser and more unstable conformation of CpxR, leading to its decreased binding affinity with downstream genes. Moreover, the Lys219 mutation resulted in the down-regulation of efflux pump genes (acrD, tolC, mdtB and mdtA), leading to the accumulation of antibiotics inside the cells and an increase in ROS production, which significantly reduces antibiotic resistance.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Conclusions<\/jats:title>\n                  <jats:p>The mutation of the key residue Lys219 causes a conformational change that results in the loss of regulatory ability of CpxR, which may potentially reduce to antibiotic resistance. Therefore, this study suggests that targeting the highly conserved sequence of CpxR could be a promising strategy for the development of new antibacterial drugs.<\/jats:p>\n               <\/jats:sec>","DOI":"10.1093\/jac\/dkad171","type":"journal-article","created":{"date-parts":[[2023,6,8]],"date-time":"2023-06-08T12:51:32Z","timestamp":1686228692000},"page":"1859-1870","source":"Crossref","is-referenced-by-count":2,"title":["Residue Lys219 of CpxR is critical in the regulation of the antibiotic resistance of <i>Escherichia coli<\/i>"],"prefix":"10.1093","volume":"78","author":[{"given":"Dan","family":"Lei","sequence":"first","affiliation":[{"name":"1MOE Key Laboratory of Tumor Molecular Biology and Key Laboratory of Functional Protein Research of Guangdong Higher Education Institutes, Institute of Life and Health Engineering, College of Life Science and Technology, Jinan University , Guangzhou , China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Linlin","family":"Cao","sequence":"additional","affiliation":[{"name":"1MOE Key Laboratory of Tumor Molecular Biology and Key Laboratory of Functional Protein Research of Guangdong Higher Education Institutes, Institute of Life and Health Engineering, College of Life Science and Technology, Jinan University , Guangzhou , China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tairan","family":"Zhong","sequence":"additional","affiliation":[{"name":"1MOE Key Laboratory of Tumor Molecular Biology and Key Laboratory of Functional Protein Research of Guangdong Higher Education Institutes, Institute of Life and Health Engineering, College of Life Science and Technology, Jinan University , Guangzhou , China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Qing-Yu","family":"He","sequence":"additional","affiliation":[{"name":"1MOE Key Laboratory of Tumor Molecular Biology and Key Laboratory of Functional Protein Research of Guangdong Higher Education Institutes, Institute of Life and Health Engineering, College of Life Science and Technology, Jinan University , Guangzhou , China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xuesong","family":"Sun","sequence":"additional","affiliation":[{"name":"1MOE Key Laboratory of Tumor Molecular Biology and Key Laboratory of Functional Protein Research of Guangdong Higher Education Institutes, Institute of Life and Health Engineering, College of Life Science and Technology, Jinan University , Guangzhou , China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"286","published-online":{"date-parts":[[2023,6,8]]},"reference":[{"key":"2023080202275877000_dkad171-B1","doi-asserted-by":"crossref","first-page":"11952","DOI":"10.1002\/anie.201504287","article-title":"An accurate in vitro model of the E. coli envelope","volume":"54","author":"Clifton","year":"2015","journal-title":"Angew Chem Int Ed 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