{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,20]],"date-time":"2026-10-20T00:53:16Z","timestamp":1792457596565,"version":"4.5.0"},"reference-count":24,"publisher":"American Society for Microbiology","issue":"2","license":[{"start":{"date-parts":[[2016,4,5]],"date-time":"2016-04-05T00:00:00Z","timestamp":1459814400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-sa\/3.0\/"},{"start":{"date-parts":[[2016,5,4]],"date-time":"2016-05-04T00:00:00Z","timestamp":1462320000000},"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":["5R01AI045626-12"],"award-info":[{"award-number":["5R01AI045626-12"]}],"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":[[2016,5,4]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            The intrinsic resistance of\n            <jats:named-content content-type=\"genus-species\">Enterococcus faecium<\/jats:named-content>\n            to ceftriaxone and cefepime (here referred to as \u201ccephalosporins\u201d) is reliant on the presence of class A penicillin-binding proteins (Pbps) PbpF and PonA. Mutants lacking these Pbps exhibit cephalosporin susceptibility that is reversible by exposure to penicillin and by selection on cephalosporin-containing medium. We selected two cephalosporin-resistant mutants (Cro1 and Cro2) of class A Pbp-deficient\n            <jats:named-content content-type=\"genus-species\">E. faecium<\/jats:named-content>\n            CV598. Genome analysis revealed changes in the serine-threonine kinase Stk in Cro1 and a truncation in the associated phosphatase StpA in Cro2 whose respective involvements in resistance were confirmed in separate complementation experiments. In an additional effort to identify proteins linked to cephalosporin resistance, we performed tandem affinity purification using Pbp5 as bait in penicillin-exposed\n            <jats:named-content content-type=\"genus-species\">E. faecium<\/jats:named-content>\n            ; these experiments yielded a protein designated Pbp5-associated protein (P\n            <jats:sub>5<\/jats:sub>\n            AP). Transcription of the P\n            <jats:sub>5<\/jats:sub>\n            AP gene was increased after exposure to penicillin in wild-type strains and in Cro2 and suppressed in Cro2 complemented with the wild-type\n            <jats:italic>stpA<\/jats:italic>\n            . Transformation of class A Pbp-deficient strains with the plasmid-carried P\n            <jats:sub>5<\/jats:sub>\n            AP gene conferred cephalosporin resistance. These data suggest that Pbp5-associated cephalosporin resistance in\n            <jats:named-content content-type=\"genus-species\">E. faecium<\/jats:named-content>\n            devoid of typical class A Pbps is related to the presence of P\n            <jats:sub>5<\/jats:sub>\n            AP, whose expression is influenced by the activity of the serine-threonine phosphatase\/kinase system.\n          <\/jats:p>\n          <jats:p>\n            <jats:bold>IMPORTANCE<\/jats:bold>\n            \u03b2-Lactam antibiotics remain our most effective therapies against susceptible Gram-positive bacteria. The intrinsic resistance of\n            <jats:named-content content-type=\"genus-species\">Enterococcus faecium<\/jats:named-content>\n            to \u03b2-lactams, particularly to cephalosporins, therefore represents a major limitation of therapy. Although the primary mechanism of resistance to \u03b2-lactams in\n            <jats:named-content content-type=\"genus-species\">E. faecium<\/jats:named-content>\n            is the presence of low-affinity monofunctional transpeptidase (class B) penicillin-binding protein Pbp5, the interaction of Pbp5 with other proteins is fundamental to maintain a resistant phenotype. The present work identifies a novel, previously uncharacterized, protein that interacts with Pbp5, whose expression increases in conjunction with stimuli that increase resistance to cephalosporins, and that confers increased resistance to cephalosporins when overexpressed. P\n            <jats:sub>5<\/jats:sub>\n            AP may represent a promising new target, inhibition of which could restore cephalosporin susceptibility to\n            <jats:named-content content-type=\"genus-species\">E. faecium<\/jats:named-content>\n            .\n          <\/jats:p>","DOI":"10.1128\/mbio.02188-15","type":"journal-article","created":{"date-parts":[[2016,4,6]],"date-time":"2016-04-06T01:13:25Z","timestamp":1459905205000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":26,"title":["Involvement of the Eukaryote-Like Kinase-Phosphatase System and a Protein That Interacts with Penicillin-Binding Protein 5 in Emergence of Cephalosporin Resistance in Cephalosporin-Sensitive Class A Penicillin-Binding Protein Mutants in Enterococcus faecium"],"prefix":"10.1128","volume":"7","author":[{"given":"Charlene","family":"Desbonnet","sequence":"first","affiliation":[{"name":"Department of Medicine, Rhode Island Hospital, Warren Alpert Medical School of Brown University, Providence, Rhode Island, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amelia","family":"Tait-Kamradt","sequence":"additional","affiliation":[{"name":"Department of Medicine, Rhode Island Hospital, Warren Alpert Medical School of Brown University, Providence, Rhode Island, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Monica","family":"Garcia-Solache","sequence":"additional","affiliation":[{"name":"Department of Medicine, Rhode Island Hospital, Warren Alpert Medical School of Brown University, Providence, Rhode Island, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paul","family":"Dunman","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, University of Rochester School of Medicine and Dentistry, Rochester, New York, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jeffrey","family":"Coleman","sequence":"additional","affiliation":[{"name":"Department of Entomology and Plant Pathology, Auburn University, Auburn, Alabama, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michel","family":"Arthur","sequence":"additional","affiliation":[{"name":"INSERM, U1138, LRMA, Equipe 12 du Centre de Recherche des Cordeliers, Paris, France"},{"name":"Universit\u00e9 Pierre et Marie Curie, UMR S 1138, Paris, France"},{"name":"Universit\u00e9 Paris Descartes, Sorbonne Paris Cit\u00e9, UMR S 1138, Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Louis B.","family":"Rice","sequence":"additional","affiliation":[{"name":"Department of Medicine, Rhode Island Hospital, Warren Alpert Medical School of Brown University, Providence, Rhode Island, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1099\/00221287-129-3-813"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.01834-08"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.45.5.1480-1486.2001"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.186.5.1221-1228.2004"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.185.24.7184-7192.2003"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.00199-11"},{"key":"e_1_3_2_8_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0608742104"},{"key":"e_1_3_2_9_2","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.01446-14"},{"key":"e_1_3_2_10_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.05984-11"},{"key":"e_1_3_2_11_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.00084-07"},{"key":"e_1_3_2_12_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2958.2009.07014.x"},{"key":"e_1_3_2_13_2","doi-asserted-by":"publisher","DOI":"10.1128\/MMBR.00042-10"},{"key":"e_1_3_2_14_2","doi-asserted-by":"publisher","DOI":"10.1038\/13732"},{"key":"e_1_3_2_15_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.01472-13"},{"key":"e_1_3_2_16_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.00859-06"},{"key":"e_1_3_2_17_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.22.2.295"},{"key":"e_1_3_2_18_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1300118110"},{"key":"e_1_3_2_19_2","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2164-14-466"},{"key":"e_1_3_2_20_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.00259-12"},{"key":"e_1_3_2_21_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0378-1097(97)00423-0"},{"key":"e_1_3_2_22_2","doi-asserted-by":"publisher","DOI":"10.1002\/pmic.200500021"},{"key":"e_1_3_2_23_2","doi-asserted-by":"publisher","DOI":"10.1006\/meth.2001.1183"},{"key":"e_1_3_2_24_2","doi-asserted-by":"publisher","DOI":"10.1093\/sysbio\/syq010"},{"key":"e_1_3_2_25_2","first-page":"2002","article-title":"A new mathematical model for relative quantification in real-time RT-PCR","volume":"29","author":"Pfaffl MW","year":"2002","unstructured":"Pfaffl MW. 2002. A new mathematical model for relative quantification in real-time RT-PCR. 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