{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,2]],"date-time":"2026-10-02T17:06:15Z","timestamp":1790960775303,"version":"4.1.0"},"reference-count":51,"publisher":"American Society for Microbiology","issue":"3","license":[{"start":{"date-parts":[[2026,3,19]],"date-time":"2026-03-19T00:00:00Z","timestamp":1773878400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,3,19]],"date-time":"2026-03-19T00:00:00Z","timestamp":1773878400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["R01 AI129541"],"award-info":[{"award-number":["R01 AI129541"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["R01 AI153160"],"award-info":[{"award-number":["R01 AI153160"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["R21 AI177818"],"award-info":[{"award-number":["R21 AI177818"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["J Bacteriol"],"published-print":{"date-parts":[[2026,3,19]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>\n                      PlrSR, a member of the NtrYX family of two-component regulatory systems (TCSs), is required for the classical bordetellae, including the causative agent of whooping cough,\n                      <jats:italic toggle=\"yes\">Bordetella pertussis<\/jats:italic>\n                      , to persist in the lower respiratory tract. The\n                      <jats:italic toggle=\"yes\">plrSR<\/jats:italic>\n                      genes are in the middle of a six-gene cluster whose regulation and roles during infection were unknown.\n                      <jats:italic toggle=\"yes\">rsmB<\/jats:italic>\n                      and\n                      <jats:italic toggle=\"yes\">plrP<\/jats:italic>\n                      are often found 5\u2032 to\n                      <jats:italic toggle=\"yes\">plrSR<\/jats:italic>\n                      homologs in \u03b2- and \u03b3-proteobacteria, while\n                      <jats:italic toggle=\"yes\">trkAH<\/jats:italic>\n                      is often found 3\u2032 to\n                      <jats:italic toggle=\"yes\">plrSR<\/jats:italic>\n                      homologs in \u237a-proteobacteria. We investigated these genes to determine if they have a functional link to\n                      <jats:italic toggle=\"yes\">plrSR<\/jats:italic>\n                      . We found that this gene cluster does not function as an operon. Rather, it contains two internal promoters: a weaker promoter in the 3\u2032 end of\n                      <jats:italic toggle=\"yes\">rsmB<\/jats:italic>\n                      and a stronger promoter in the 3\u2032 end of\n                      <jats:italic toggle=\"yes\">plrS<\/jats:italic>\n                      . Additionally, our results indicate that PlrP functions as a third component of the PlrSR TCS. Genetic manipulations of\n                      <jats:italic toggle=\"yes\">plrP, plrS,<\/jats:italic>\n                      and\n                      <jats:italic toggle=\"yes\">plrR<\/jats:italic>\n                      indicate that PlrP is essential\n                      <jats:italic toggle=\"yes\">in vitro<\/jats:italic>\n                      and strongly suggest that it inhibits PlrS phosphatase activity, likely through PlrS\u2019s PhoQ-DcuS-CitA (PDC) domain. Since our results indicate that PlrR can be phosphorylated by another unknown phosphodonor\n                      <jats:italic toggle=\"yes\">in vitro<\/jats:italic>\n                      , limiting PlrS phosphatase activity ensures PlrR~P is not dephosphorylated to lethally low levels. Using natural-host models, we determined that high levels of PlrR~P are required for bacterial survival in the lower respiratory tract, and that PlrP affects PlrS activity\n                      <jats:italic toggle=\"yes\">in vivo<\/jats:italic>\n                      . Given that\n                      <jats:italic toggle=\"yes\">plrP<\/jats:italic>\n                      homologs always colocalize with\n                      <jats:italic toggle=\"yes\">ntrYX<\/jats:italic>\n                      homologs, we propose that PlrP may fulfill similar functions in other \u03b2- and \u03b3-proteobacteria that encode NtrYX-family TCSs, including nonpathogens.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>IMPORTANCE<\/jats:title>\n                    <jats:p>\n                      <jats:italic toggle=\"yes\">Bordetella<\/jats:italic>\n                      species, including\n                      <jats:italic toggle=\"yes\">B. pertussis<\/jats:italic>\n                      , cause respiratory infections in humans and other animals. Their PlrSR two-component regulatory systems are required for survival in the lower respiratory tract. We characterized the six-gene cluster that includes\n                      <jats:italic toggle=\"yes\">plrS<\/jats:italic>\n                      and\n                      <jats:italic toggle=\"yes\">plrR<\/jats:italic>\n                      , identifying two internal promoters that drive expression of\n                      <jats:italic toggle=\"yes\">plrP<\/jats:italic>\n                      and\n                      <jats:italic toggle=\"yes\">plrS<\/jats:italic>\n                      separately from\n                      <jats:italic toggle=\"yes\">plrR<\/jats:italic>\n                      . Our data indicate that the\n                      <jats:italic toggle=\"yes\">plrP<\/jats:italic>\n                      gene product is an essential third component of the PlrSR two-component regulatory system (TCS), functioning to prevent PlrS from acting as a strong phosphatase\n                      <jats:italic toggle=\"yes\">in vitro<\/jats:italic>\n                      . PlrP homologs are present, and may function similarly, in NtrYX-family TCSs in other \u03b2- and \u03b3-proteobacteria. Our results are important because they provide insight into how bacteria, including pathogens, sense and respond to their environment.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1128\/jb.00521-25","type":"journal-article","created":{"date-parts":[[2026,2,13]],"date-time":"2026-02-13T14:00:25Z","timestamp":1770991225000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":1,"title":["Genetic evidence for a periplasmic protein as a third component for a subset of NtrYX family two-component systems"],"prefix":"10.1128","volume":"208","author":[{"ORCID":"https:\/\/orcid.org\/0009-0003-7775-8039","authenticated-orcid":true,"given":"Alexa R.","family":"Wolber","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/0130frc33","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Microbiology and Immunology, School of Medicine, University of North Carolina-Chapel Hill","place":["Chapel Hill, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5426-5539","authenticated-orcid":true,"given":"Liliana S.","family":"McKay","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/0130frc33","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Microbiology and Immunology, School of Medicine, University of North Carolina-Chapel Hill","place":["Chapel Hill, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Richard M.","family":"Johnson","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/0130frc33","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Microbiology and Immunology, School of Medicine, University of North Carolina-Chapel Hill","place":["Chapel Hill, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zain T.","family":"Hameed","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/0130frc33","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Microbiology and Immunology, School of Medicine, University of North Carolina-Chapel Hill","place":["Chapel Hill, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Katlyn B.","family":"Mote","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/0130frc33","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Microbiology and Immunology, School of Medicine, University of North Carolina-Chapel Hill","place":["Chapel Hill, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Steven M.","family":"Julio","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00xhcz327","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Biology, Westmont College","place":["Santa Barbara, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3726-3990","authenticated-orcid":true,"given":"Peggy A.","family":"Cotter","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/0130frc33","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Microbiology and Immunology, School of Medicine, University of North Carolina-Chapel Hill","place":["Chapel Hill, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_4_2_2","doi-asserted-by":"publisher","DOI":"10.1128\/aem.57.4.1202-1206.1991"},{"key":"e_1_3_4_3_2","unstructured":"CDC. 2025. 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