{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T16:00:00Z","timestamp":1791302400972,"version":"4.1.0"},"reference-count":26,"publisher":"American Chemical Society (ACS)","issue":"39","license":[{"start":{"date-parts":[[2026,9,27]],"date-time":"2026-09-27T00:00:00Z","timestamp":1790467200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/100031206","name":"Yamada Science Foundation","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100031206","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100008655","name":"Hamaguchi Foundation for the Advancement of Biochemistry","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100008655","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"publisher","award":["26K08125"],"award-info":[{"award-number":["26K08125"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100009666","name":"Salt Science Research Foundation","doi-asserted-by":"publisher","award":["202605"],"award-info":[{"award-number":["202605"]}],"id":[{"id":"10.13039\/100009666","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,10,6]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Arsenite [As(III)]-responsive transcriptional regulation is widely employed by microorganisms for detoxification and homeostasis. Engineering As(III)-responsive behavior into transcription factors that are intrinsically metal-insensitive remains challenging. Here, we introduced a third cysteine residue at different positions in the engineered BetIL79C\/Q81C scaffold and evaluated the resulting whole-cell sfGFP reporter responses. The magnitude and direction of the As(III)-dependent reporter change varied with the position of the introduced cysteine. Under combined As(III) and choline inputs, selected variants exhibited operational OR-like or As(III)-inhibited reporter-output patterns over defined concentration ranges. To explore possible structural interpretations for these reporter phenotypes, we performed molecular dynamics simulations using predefined dithiol- and trithiol-coordination models. Simulations of these models yielded distinct fluctuation patterns in the DNA-binding domain, providing testable structural hypotheses for the observed reporter phenotypes. Because neither As(III) binding to the engineered cysteine sites nor As(III)-dependent changes in BetI\u2013DNA binding were directly measured, the reporter assays and simulations do not establish the underlying molecular mechanism. Nevertheless, our experiments show that cysteine placement can tune the direction of As(III)-dependent whole-cell sfGFP output in an engineered BetI regulatory system and thereby expand the design space for multi-input transcriptional devices.<\/jats:p>","DOI":"10.1021\/acsomega.6c04287","type":"journal-article","created":{"date-parts":[[2026,9,27]],"date-time":"2026-09-27T15:01:26Z","timestamp":1790521286000},"page":"59054-59063","source":"Crossref","is-referenced-by-count":0,"title":["Position-Dependent\nActivation and Repression by As(III)\nin an Engineered BetI Transcriptional Repressor"],"prefix":"10.1021","volume":"11","author":[{"given":"Katsumasa","family":"Kamiya","sequence":"first","affiliation":[{"name":"Kanagawa Institute of Technology , , , ,","place":["Atsugi, Kanagawa, Japan, 243-0292"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ryo","family":"Yamaguchi","sequence":"additional","affiliation":[{"name":"Chiba University , , ,","place":["Chiba, Japan, 263-8522"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kojiro","family":"Shimizu","sequence":"additional","affiliation":[{"name":"Chiba University , , ,","place":["Chiba, Japan, 263-8522"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4753-7652","authenticated-orcid":true,"given":"Shigeko","family":"Kawai-Noma","sequence":"additional","affiliation":[{"name":"Chiba University , , ,","place":["Chiba, Japan, 263-8522"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,9,27]]},"reference":[{"key":"2026100610470606000_cit1","volume-title":"Arsenic and Arsenic Compounds","author":"Ng","year":"2001","edition":"2nd"},{"issue":"2\u20133","key":"2026100610470606000_cit2","doi-asserted-by":"publisher","first-page":"131","DOI":"10.1016\/S0168-6445(03)00054-8","article-title":"The SmtB\/ArsR Family\nof Metalloregulatory\nTranscriptional Repressors: Structural Insights into Prokaryotic Metal\nResistance","volume":"27","author":"Busenlehner","year":"2003","journal-title":"FEMS Microbiol. Rev."},{"issue":"43","key":"2026100610470606000_cit3","doi-asserted-by":"publisher","first-page":"23530","DOI":"10.1021\/acs.est.5c11929","article-title":"Evolutionary Advantages of Multiple Arsenic Binding\nSites in an ArsR Transcriptional Repressor","volume":"59","author":"Chen","year":"2025","journal-title":"Environ. Sci. Technol."},{"key":"2026100610470606000_cit4","doi-asserted-by":"publisher","first-page":"19","DOI":"10.1002\/9780470975503.ch2","article-title":"Arsenic\u2019s\nInteractions with Macromolecules and Its Relationship to Carcinogenesis","volume-title":"Biological Chemistry of Arsenic, Antimony and Bismuth","author":"Kitchin","year":"2010"},{"issue":"14","key":"2026100610470606000_cit5","doi-asserted-by":"publisher","first-page":"16035","DOI":"10.1021\/acsomega.3c09604","article-title":"Imparting\nAs(III) Responsiveness to the Choline Response Transcriptional Regulator\nBetI","volume":"9","author":"Yamaguchi","year":"2024","journal-title":"ACS Omega"},{"issue":"4","key":"2026100610470606000_cit6","doi-asserted-by":"publisher","first-page":"491","DOI":"10.1093\/bbb\/zbaf193","article-title":"As(III)-Induced Protein\nAggregation: Insights and Emerging Possibilities for As(III)-Responsive\nProteins","volume":"90","author":"Kawai-Noma","year":"2026","journal-title":"Biosci., Biotechnol., Biochem."},{"issue":"10","key":"2026100610470606000_cit7","doi-asserted-by":"publisher","first-page":"7769","DOI":"10.1021\/cr300015c","article-title":"Arsenic\nBinding to Proteins","volume":"113","author":"Shen","year":"2013","journal-title":"Chem. Rev."},{"issue":"2","key":"2026100610470606000_cit8","doi-asserted-by":"publisher","first-page":"P25","DOI":"10.1016\/j.ccell.2020.11.013","article-title":"Arsenic Trioxide Rescues\nStructural P53 Mutations through\na Cryptic Allosteric Site","volume":"39","author":"Chen","year":"2021","journal-title":"Cancer Cell"},{"issue":"2","key":"2026100610470606000_cit9","doi-asserted-by":"publisher","first-page":"326","DOI":"10.1128\/MMBR.69.2.326-356.2005","article-title":"The TetR Family of\nTranscriptional\nRepressors","volume":"69","author":"Ramos","year":"2005","journal-title":"Microbiol. Mol. Biol. Rev."},{"issue":"6","key":"2026100610470606000_cit10","doi-asserted-by":"publisher","first-page":"1663","DOI":"10.1128\/jb.178.6.1663-1670.1996","article-title":"DNA-Binding Properties of the BetI\nRepressor Protein of Escherichia Coli: The Inducer Choline Stimulates\nBetI-DNA Complex Formation","volume":"178","author":"Rkenes","year":"1996","journal-title":"J. Bacteriol."},{"issue":"6","key":"2026100610470606000_cit11","doi-asserted-by":"publisher","first-page":"1655","DOI":"10.1128\/jb.178.6.1655-1662.1996","article-title":"The Complex\nBet Promoters of Escherichia Coli: Regulation by Oxygen (ArcA), Choline\n(BetI), and Osmotic Stress","volume":"178","author":"Lamark","year":"1996","journal-title":"J. Bacteriol."},{"issue":"12","key":"2026100610470606000_cit12","doi-asserted-by":"publisher","first-page":"1352","DOI":"10.1021\/acssynbio.5b00107","article-title":"Evolutionary Design of Choline-Inducible\nand -Repressible T7-Based Induction Systems","volume":"4","author":"Ike","year":"2015","journal-title":"ACS Synth. 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