{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T09:16:15Z","timestamp":1785402975056,"version":"3.56.0"},"reference-count":37,"publisher":"American Society for Microbiology","issue":"6","license":[{"start":{"date-parts":[[2026,6,18]],"date-time":"2026-06-18T00:00:00Z","timestamp":1781740800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,6,18]],"date-time":"2026-06-18T00:00:00Z","timestamp":1781740800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"publisher","award":["20K07428, 23K06529"],"award-info":[{"award-number":["20K07428, 23K06529"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"publisher","award":["23K14521, 25K18800"],"award-info":[{"award-number":["23K14521, 25K18800"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"publisher","award":["23K05637"],"award-info":[{"award-number":["23K05637"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100031489","name":"Shionogi Infectious Disease Research Promotion Foundation","doi-asserted-by":"crossref","award":["2024N068"],"award-info":[{"award-number":["2024N068"]}],"id":[{"id":"10.13039\/100031489","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100019670","name":"BIKEN Foundation","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100019670","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["J Bacteriol"],"published-print":{"date-parts":[[2026,6,18]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>\n                      <jats:italic toggle=\"yes\">Vibrio parahaemolyticus<\/jats:italic>\n                      , a gram-negative marine bacterium, is a major cause of seafood-borne gastroenteritis worldwide. This pathogen relies on type III secretion system 2 (T3SS2), which is encoded on a pathogenicity island, for its enteropathogenicity. Expression of T3SS2 is activated by a regulatory pathway centered on the transcriptional activator VtrB, which is antagonized by the xenogeneic silencer, histone-like nucleoid-structuring protein (H-NS). However, the complete transcriptional network is not yet fully understood. In this study, we identified TsrA as a negative regulator of T3SS2 gene expression. TsrA is a small protein conserved among\n                      <jats:italic toggle=\"yes\">Vibrio<\/jats:italic>\n                      species that lacks a putative DNA-binding motif but has been implicated in the regulation of virulence genes in\n                      <jats:italic toggle=\"yes\">Vibrio cholerae<\/jats:italic>\n                      . In\n                      <jats:italic toggle=\"yes\">V. parahaemolyticus<\/jats:italic>\n                      , deletion of\n                      <jats:italic toggle=\"yes\">tsrA<\/jats:italic>\n                      increased VtrB production and T3SS2 secretion, thereby enhancing T3SS2-dependent pathogenicity. Transcription of\n                      <jats:italic toggle=\"yes\">vtrB<\/jats:italic>\n                      occurs via a two-step activation process, in which TsrA affects the primary activation step, thereby modulating VtrB production. We further provide experimental evidence that TsrA physically interacts with H-NS via its C-terminal region, which correlates with its regulatory activity on\n                      <jats:italic toggle=\"yes\">vtrB<\/jats:italic>\n                      expression. A systematic mutational analysis of the C-terminal 26 residues revealed several residues critical for TsrA regulatory activity. Moreover, the regulatory effect of TsrA on T3SS2 gene expression was dependent on H-NS, demonstrating that TsrA functions in concert with H-NS. Thus, our findings provide new insights into the regulatory mechanisms of virulence gene expression in\n                      <jats:italic toggle=\"yes\">V. parahaemolyticus<\/jats:italic>\n                      by defining the role of TsrA in this network, while also placing TsrA among H-NS co-regulators.\n                    <\/jats:p>\n                    <jats:sec>\n                      <jats:title>IMPORTANCE<\/jats:title>\n                      <jats:p>\n                        Nucleoid-associated proteins (NAPs) play key roles in virulence gene regulation in bacteria. The best-studied NAP is H-NS, which often functions with co-regulators to fine-tune gene expression. TsrA, a small protein lacking a DNA-binding motif conserved among\n                        <jats:italic toggle=\"yes\">Vibrio<\/jats:italic>\n                        species, has been suggested to be functionally related to H-NS in\n                        <jats:italic toggle=\"yes\">Vibrio cholerae<\/jats:italic>\n                        , although its mechanism remains unknown. Here, we demonstrate that TsrA negatively regulates the expression of type III secretion system 2 (T3SS2), a major virulence determinant of\n                        <jats:italic toggle=\"yes\">Vibrio parahaemolyticus<\/jats:italic>\n                        , an important seafood-borne pathogen. TsrA modulates the transcription of\n                        <jats:italic toggle=\"yes\">vtrB<\/jats:italic>\n                        , which encodes the essential activator for T3SS2 expression, through direct physical interaction with H-NS. Our findings reveal a molecular link between TsrA and H-NS, providing mechanistic insights into NAP- and TsrA-mediated regulation of virulence in\n                        <jats:italic toggle=\"yes\">Vibrio<\/jats:italic>\n                        .\n                      <\/jats:p>\n                    <\/jats:sec>\n                  <\/jats:sec>","DOI":"10.1128\/jb.00556-25","type":"journal-article","created":{"date-parts":[[2026,5,28]],"date-time":"2026-05-28T14:20:00Z","timestamp":1779978000000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":0,"title":["TsrA modulates type III secretion system 2 expression as a co-regulator of H-NS in\n                    <i>Vibrio parahaemolyticus<\/i>"],"prefix":"10.1128","volume":"208","author":[{"given":"Tan Paramita Wibowo","family":"Sutanto","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/035t8zc32","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Bacterial Infections, Research Institute for Microbial Diseases, The University of Osaka","place":["Osaka, Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5384-4475","authenticated-orcid":true,"given":"Andre","family":"Pratama","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/035t8zc32","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Bacterial Infections, Research Institute for Microbial Diseases, The University of Osaka","place":["Osaka, Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7899-252X","authenticated-orcid":false,"given":"Eiji","family":"Ishii","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/035t8zc32","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Bacterial Infections, Research Institute for Microbial Diseases, The University of Osaka","place":["Osaka, Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5134-1794","authenticated-orcid":true,"given":"Tetsuya","family":"Iida","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/035t8zc32","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Bacterial Infections, Research Institute for Microbial Diseases, The University of Osaka","place":["Osaka, Japan"]},{"id":[{"id":"https:\/\/ror.org\/035t8zc32","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Infection Metagenomics, Bioinformatics Center, Research Institute for Microbial Diseases, The University of Osaka","place":["Osaka, Japan"]},{"id":[{"id":"https:\/\/ror.org\/035t8zc32","id-type":"ROR","asserted-by":"publisher"}],"name":"Center for Infectious Disease Education and Research, The University of Osaka","place":["Osaka, Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8430-9723","authenticated-orcid":true,"given":"Shigeaki","family":"Matsuda","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/035t8zc32","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Bacterial Infections, Research Institute for Microbial Diseases, The University of Osaka","place":["Osaka, Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_5_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.tim.2020.02.008"},{"key":"e_1_3_5_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/S2542-5196(24)00124-4"},{"key":"e_1_3_5_4_2","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.00013-07"},{"key":"e_1_3_5_5_2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-micro-092412-155725"},{"key":"e_1_3_5_6_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.00266-22"},{"key":"e_1_3_5_7_2","doi-asserted-by":"publisher","DOI":"10.1038\/nrmicro.2017.20"},{"key":"e_1_3_5_8_2","doi-asserted-by":"publisher","DOI":"10.1111\/1348-0421.12767"},{"key":"e_1_3_5_9_2","doi-asserted-by":"publisher","DOI":"10.1128\/mSphere.00599-21"},{"key":"e_1_3_5_10_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2023.113261"},{"key":"e_1_3_5_11_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0008678"},{"key":"e_1_3_5_12_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0187846"},{"key":"e_1_3_5_13_2","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.15718"},{"key":"e_1_3_5_14_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1012094"},{"key":"e_1_3_5_15_2","doi-asserted-by":"publisher","DOI":"10.1128\/mSphere.01014-20"},{"key":"e_1_3_5_16_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1014998107"},{"key":"e_1_3_5_17_2","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.02901-20"},{"key":"e_1_3_5_18_2","doi-asserted-by":"publisher","DOI":"10.1128\/msphere.00011-24"},{"key":"e_1_3_5_19_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.01051-09"},{"key":"e_1_3_5_20_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0013365"},{"key":"e_1_3_5_21_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M113.455378"},{"key":"e_1_3_5_22_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gky265"},{"key":"e_1_3_5_23_2","doi-asserted-by":"publisher","DOI":"10.1038\/srep00509"},{"key":"e_1_3_5_24_2","doi-asserted-by":"publisher","DOI":"10.1111\/mmi.15283"},{"key":"e_1_3_5_25_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.178.15.4335-4343.1996"},{"key":"e_1_3_5_26_2","doi-asserted-by":"publisher","DOI":"10.1007\/s004380051178"},{"key":"e_1_3_5_27_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2006.03.059"},{"key":"e_1_3_5_28_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.01124-06"},{"key":"e_1_3_5_29_2","doi-asserted-by":"publisher","DOI":"10.1038\/nrmicro883"},{"key":"e_1_3_5_30_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(03)12659-1"},{"key":"e_1_3_5_31_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.72.11.6659-6665.2004"},{"key":"e_1_3_5_32_2","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.1923"},{"key":"e_1_3_5_33_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41564-019-0368-y"},{"key":"e_1_3_5_34_2","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/btu638"},{"key":"e_1_3_5_35_2","doi-asserted-by":"publisher","DOI":"10.1186\/s12859-018-2486-6"},{"key":"e_1_3_5_36_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(19)68491-7"},{"key":"e_1_3_5_37_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-024-07487-w"},{"key":"e_1_3_5_38_2","doi-asserted-by":"publisher","DOI":"10.1002\/pro.4792"}],"container-title":["Journal of Bacteriology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/jb.00556-25","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/jb.00556-25","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,3]],"date-time":"2026-07-03T13:10:23Z","timestamp":1783084223000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/jb.00556-25"}},"subtitle":[],"editor":[{"given":"Tao","family":"Dong","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2026,6,18]]},"references-count":37,"journal-issue":{"issue":"6","published-print":{"date-parts":[[2026,6,18]]}},"alternative-id":["10.1128\/jb.00556-25"],"URL":"https:\/\/doi.org\/10.1128\/jb.00556-25","relation":{},"ISSN":["0021-9193","1098-5530"],"issn-type":[{"value":"0021-9193","type":"print"},{"value":"1098-5530","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,6,18]]},"assertion":[{"value":"2025-11-26","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-04-07","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-05-28","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e00556-25"}}