{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,22]],"date-time":"2026-10-22T05:39:00Z","timestamp":1792647540394,"version":"4.5.0"},"reference-count":39,"publisher":"American Chemical Society (ACS)","issue":"1","license":[{"start":{"date-parts":[[2019,10,14]],"date-time":"2019-10-14T00:00:00Z","timestamp":1571011200000},"content-version":"vor","delay-in-days":18,"URL":"https:\/\/pubs.acs.org\/page\/policy\/authorchoice_ccby_termsofuse.html"}],"funder":[{"DOI":"10.13039\/100000060","name":"National Institute of Allergy and Infectious Diseases","doi-asserted-by":"publisher","award":["R01AI116605"],"award-info":[{"award-number":["R01AI116605"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000060","name":"National Institute of Allergy and Infectious Diseases","doi-asserted-by":"publisher","award":["R21AI105687"],"award-info":[{"award-number":["R21AI105687"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000865","name":"Bill and Melinda Gates Foundation","doi-asserted-by":"publisher","award":["OPP1059227"],"award-info":[{"award-number":["OPP1059227"]}],"id":[{"id":"10.13039\/100000865","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000865","name":"Bill and Melinda Gates Foundation","doi-asserted-by":"publisher","award":["OPP1119065"],"award-info":[{"award-number":["OPP1119065"]}],"id":[{"id":"10.13039\/100000865","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100007709","name":"Michigan State University","doi-asserted-by":"publisher","award":["NA"],"award-info":[{"award-number":["NA"]}],"id":[{"id":"10.13039\/100007709","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100011138","name":"AgBioResearch, Michigan State University","doi-asserted-by":"publisher","award":["NA"],"award-info":[{"award-number":["NA"]}],"id":[{"id":"10.13039\/100011138","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2020,1,17]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Mycobacterium tuberculosis (Mtb) possesses a two-component regulatory system, DosRST, that enables Mtb to sense host immune cues and establish a state of nonreplicating persistence (NRP). NRP bacteria are tolerant to several antimycobacterial drugs in vitro and are thought to play a role in the long course of tuberculosis therapy. Previously, we reported the discovery of six novel chemical inhibitors of DosRST, named HC101A\u2013106A, from a whole cell, reporter-based phenotypic high throughput screen. Here, we report functional and mechanism of action studies of HC104A and HC106A. RNaseq transcriptional profiling shows that the compounds downregulate genes of the DosRST regulon. Both compounds reduce hypoxia-induced triacylglycerol synthesis by \u223c50%. HC106A inhibits Mtb survival during hypoxia-induced NRP; however, HC104A did not inhibit survival during NRP. An electrophoretic mobility assay shows that HC104A inhibits DosR DNA binding in a dose-dependent manner, indicating that HC104A may function by directly targeting DosR. In contrast, UV\u2013visible spectroscopy studies suggest HC106A directly targets the sensor kinase heme, via a mechanism that is distinct from the oxidation and alkylation of heme previously observed with artemisinin (HC101A). Synergistic interactions were observed when DosRST inhibitors were examined in pairwise combinations with the strongest potentiation observed between artemisinin paired with HC102A, HC103A, or HC106A. Our data collectively show that the DosRST pathway can be inhibited by multiple distinct mechanisms.<\/jats:p>","DOI":"10.1021\/acschembio.8b00849","type":"journal-article","created":{"date-parts":[[2019,9,26]],"date-time":"2019-09-26T12:00:37Z","timestamp":1569499237000},"page":"52-62","source":"Crossref","is-referenced-by-count":62,"title":["Inhibiting\n                    <i>Mycobacterium tuberculosis<\/i>\n                    DosRST Signaling by Targeting Response Regulator DNA Binding and\nSensor Kinase Heme"],"prefix":"10.1021","volume":"15","author":[{"given":"Huiqing","family":"Zheng","sequence":"first","affiliation":[{"name":"Michigan State University , , , ,","place":["East Lansing, Michigan, United States, 48824"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"John T.","family":"Williams","sequence":"additional","affiliation":[{"name":"Michigan State University , , , ,","place":["East Lansing, Michigan, United States, 48824"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bilal","family":"Aleiwi","sequence":"additional","affiliation":[{"name":"Michigan State University , , , ,","place":["East Lansing, Michigan, United States, 48824"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Edmund","family":"Ellsworth","sequence":"additional","affiliation":[{"name":"Michigan State University , , , ,","place":["East Lansing, Michigan, United States, 48824"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4119-4169","authenticated-orcid":true,"given":"Robert B.","family":"Abramovitch","sequence":"additional","affiliation":[{"name":"Michigan State University , , , ,","place":["East Lansing, Michigan, United States, 48824"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2019,9,26]]},"reference":[{"key":"2026090213591904500_cit1","doi-asserted-by":"publisher","first-page":"1151","DOI":"10.1111\/j.1462-5822.2009.01325.x","article-title":"Hypoxia: a window\ninto Mycobacterium tuberculosis latency","volume":"11","author":"Rustad","year":"2009","journal-title":"Cell.\nMicrobiol."},{"key":"2026090213591904500_cit2","doi-asserted-by":"publisher","first-page":"139","DOI":"10.1146\/annurev.micro.55.1.139","article-title":"Nonreplicating persistence of mycobacterium tuberculosis","volume":"55","author":"Wayne","year":"2001","journal-title":"Annu. Rev. Microbiol."},{"key":"2026090213591904500_cit3","doi-asserted-by":"publisher","first-page":"1185","DOI":"10.1164\/rccm.201408-1502OC","article-title":"The DosR Regulon\nModulates Adaptive Immunity and Is\nEssential for Mycobacterium tuberculosis Persistence","volume":"191","author":"Mehra","year":"2015","journal-title":"Am. J. Respir. Crit. Care Med."},{"key":"2026090213591904500_cit4","doi-asserted-by":"publisher","first-page":"637","DOI":"10.1165\/rcmb.2016-0239OC","article-title":"Hypoxia Sensing\nand Persistence Genes Are Expressed during the Intragranulomatous\nSurvival of Mycobacterium tuberculosis","volume":"56","author":"Hudock","year":"2017","journal-title":"Am.\nJ. Respir. Cell Mol. Biol."},{"key":"2026090213591904500_cit5","doi-asserted-by":"publisher","first-page":"141","DOI":"10.1054\/tuld.2000.0240","article-title":"Characterization\nof a two-component\nsystem, devR-devS, of Mycobacterium tuberculosis","volume":"80","author":"Dasgupta","year":"2000","journal-title":"Tubercle and lung disease: the official journal of the International\nUnion against Tuber Lung Dis"},{"key":"2026090213591904500_cit6","doi-asserted-by":"publisher","first-page":"833","DOI":"10.1046\/j.1365-2958.2003.03474.x","article-title":"Rv3133c\/dosR is\na transcription factor that mediates the hypoxic response of Mycobacterium\ntuberculosis","volume":"48","author":"Park","year":"2003","journal-title":"Mol. Microbiol."},{"key":"2026090213591904500_cit7","doi-asserted-by":"publisher","first-page":"865","DOI":"10.1099\/mic.0.26218-0","article-title":"DevR-DevS is a bona\nfide two-component system of Mycobacterium tuberculosis that is hypoxia-responsive in\nthe absence of the DNA-binding domain of DevR","volume":"150","author":"Saini","year":"2004","journal-title":"Microbiology (London, U. K.)"},{"key":"2026090213591904500_cit8","doi-asserted-by":"publisher","first-page":"705","DOI":"10.1084\/jem.20030205","article-title":"Inhibition of respiration by nitric\noxide induces a Mycobacterium tuberculosis dormancy program","volume":"198","author":"Voskuil","year":"2003","journal-title":"J. Exp. Med."},{"key":"2026090213591904500_cit9","doi-asserted-by":"publisher","first-page":"18032","DOI":"10.1074\/jbc.M802274200","article-title":"Heme oxygenase-1-derived\ncarbon monoxide induces the Mycobacterium tuberculosis dormancy regulon","volume":"283","author":"Kumar","year":"2008","journal-title":"J. Biol. Chem."},{"key":"2026090213591904500_cit10","doi-asserted-by":"publisher","first-page":"6795","DOI":"10.1128\/JB.00401-08","article-title":"O2- and NO-sensing mechanism through\nthe DevSR two-component system in Mycobacterium smegmatis","volume":"190","author":"Lee","year":"2008","journal-title":"Journal of bacteriology"},{"key":"2026090213591904500_cit11","doi-asserted-by":"publisher","first-page":"1708","DOI":"10.1110\/ps.072897707","article-title":"DosT and\nDevS are oxygen-switched kinases in Mycobacterium tuberculosis","volume":"16","author":"Sousa","year":"2007","journal-title":"Protein Sci."},{"key":"2026090213591904500_cit12","doi-asserted-by":"publisher","first-page":"11568","DOI":"10.1073\/pnas.0705054104","article-title":"Mycobacterium tuberculosis\nDosS is\na redox sensor and DosT is a hypoxia sensor","volume":"104","author":"Kumar","year":"2007","journal-title":"Proc. Natl. Acad. Sci. U. S. A."},{"key":"2026090213591904500_cit13","doi-asserted-by":"publisher","first-page":"4250","DOI":"10.1021\/bi602422p","article-title":"DevS, a\nheme-containing two-component oxygen sensor of Mycobacterium tuberculosis","volume":"46","author":"Ioanoviciu","year":"2007","journal-title":"Biochemistry"},{"key":"2026090213591904500_cit14","doi-asserted-by":"publisher","first-page":"4868","DOI":"10.1128\/JB.00550-10","article-title":"Different\nroles of DosS and DosT in the hypoxic adaptation of Mycobacteria","volume":"192","author":"Kim","year":"2010","journal-title":"J. Bacteriol."},{"key":"2026090213591904500_cit15","doi-asserted-by":"publisher","first-page":"11945","DOI":"10.1073\/pnas.0711697105","article-title":"The protonmotive\nforce is required for maintaining ATP homeostasis and viability of\nhypoxic, nonreplicating Mycobacterium tuberculosis","volume":"105","author":"Rao","year":"2008","journal-title":"Proc. Natl. Acad. Sci. U. S. A."},{"key":"2026090213591904500_cit16","doi-asserted-by":"publisher","first-page":"39","DOI":"10.1016\/j.cell.2011.02.022","article-title":"Drug tolerance in replicating mycobacteria\nmediated by a macrophage-induced efflux mechanism","volume":"145","author":"Adams","year":"2011","journal-title":"Cell"},{"key":"2026090213591904500_cit17","doi-asserted-by":"publisher","first-page":"12147","DOI":"10.1073\/pnas.1203735109","article-title":"Eradication\nof bacterial persisters with antibiotic-generated hydroxyl radicals","volume":"109","author":"Grant","year":"2012","journal-title":"Proc. Natl. Acad. Sci. U. S. A."},{"key":"2026090213591904500_cit18","doi-asserted-by":"publisher","first-page":"1662","DOI":"10.1128\/JB.00926-09","article-title":"The Mycobacterium\ntuberculosis DosR regulon assists\nin metabolic homeostasis and enables rapid recovery from nonrespiring\ndormancy","volume":"192","author":"Leistikow","year":"2010","journal-title":"J. Bacteriol."},{"key":"2026090213591904500_cit19","doi-asserted-by":"publisher","first-page":"1230","DOI":"10.1128\/IAI.01117-08","article-title":"Role of the dosR-dosS two-component regulatory system\nin Mycobacterium tuberculosis virulence in three animal models","volume":"77","author":"Converse","year":"2009","journal-title":"Infect. Immun."},{"key":"2026090213591904500_cit20","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0135208","article-title":"in-vivo gene signatures\nof Mycobacterium tuberculosis in C3HeB\/FeJ Mice","volume":"10","author":"Gautam","year":"2015","journal-title":"PLoS One"},{"key":"2026090213591904500_cit21","doi-asserted-by":"publisher","first-page":"708","DOI":"10.1165\/rcmb.2014-0230OC","article-title":"DosS is required for\nthe complete virulence of Mycobacterium tuberculosis in mice with classical granulomatous lesions","volume":"52","author":"Gautam","year":"2015","journal-title":"Am. J. Respir. Cell Mol. Biol."},{"key":"2026090213591904500_cit22","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0006077","article-title":"A novel\nin vitro multiple-stress dormancy model for Mycobacterium tuberculosis\ngenerates a lipid-loaded, drug-tolerant, dormant pathogen","volume":"4","author":"Deb","year":"2009","journal-title":"PLoS One"},{"key":"2026090213591904500_cit23","doi-asserted-by":"publisher","first-page":"5503","DOI":"10.1039\/C7OB01028A","article-title":"Antimicrobial peptide-inspired NH125\nanalogues: bacterial\nand fungal biofilm-eradicating agents and rapid killers of MRSA persisters","volume":"15","author":"Basak","year":"2017","journal-title":"Org. Biomol. Chem."},{"key":"2026090213591904500_cit24","doi-asserted-by":"publisher","first-page":"339","DOI":"10.1016\/j.tips.2017.01.004","article-title":"Small Molecules That Sabotage Bacterial Virulence","volume":"38","author":"Johnson","year":"2017","journal-title":"Trends Pharmacol. Sci."},{"key":"2026090213591904500_cit25","doi-asserted-by":"publisher","first-page":"218","DOI":"10.1038\/nchembio.2259","article-title":"Inhibitors\nof Mycobacterium tuberculosis DosRST signaling and persistence","volume":"13","author":"Zheng","year":"2017","journal-title":"Nat. Chem. Biol."},{"key":"2026090213591904500_cit26","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pbio.1001065","article-title":"Metabolic regulation of mycobacterial\ngrowth and antibiotic sensitivity","volume":"9","author":"Baek","year":"2011","journal-title":"PLoS Biol."},{"key":"2026090213591904500_cit27","doi-asserted-by":"publisher","first-page":"1190","DOI":"10.1021\/cb2004884","article-title":"A high-throughput screen to identify inhibitors of\nATP homeostasis in non-replicating Mycobacterium tuberculosis","volume":"7","author":"Mak","year":"2012","journal-title":"ACS Chem. Biol."},{"key":"2026090213591904500_cit28","doi-asserted-by":"publisher","first-page":"670","DOI":"10.1126\/science.1116739","article-title":"Small-molecule\ninhibitor of Vibrio cholerae virulence and intestinal colonization","volume":"310","author":"Hung","year":"2005","journal-title":"Science"},{"key":"2026090213591904500_cit29","doi-asserted-by":"publisher","first-page":"12523","DOI":"10.1021\/bi8012356","article-title":"2.3 A X-ray structure\nof the heme-bound GAF domain of sensory histidine kinase DosT of Mycobacterium\ntuberculosis","volume":"47","author":"Podust","year":"2008","journal-title":"Biochemistry"},{"key":"2026090213591904500_cit30","doi-asserted-by":"publisher","first-page":"13057","DOI":"10.1074\/jbc.M808905200","article-title":"Structural\ninsight into the heme-based redox sensing by DosS from Mycobacterium\ntuberculosis","volume":"284","author":"Cho","year":"2009","journal-title":"J. Biol. Chem."},{"key":"2026090213591904500_cit31","unstructured":"Chou, T., and Martin, N. (2005)  CompuSyn\nfor drug combinations: PC software and user\u2019s\nguide: a computer program for quantitation of synergism and antagonism\nin drug combinations, and the determination of IC50 and ED50 and LD50\nvalues, ComboSyn, Paramus, NJ."},{"key":"2026090213591904500_cit32","doi-asserted-by":"publisher","first-page":"27","DOI":"10.1016\/0065-2571(84)90007-4","article-title":"Quantitative analysis of dose-effect relationships:\nthe combined effects of multiple drugs or enzyme inhibitors","volume":"22","author":"Chou","year":"1984","journal-title":"Adv. Enzyme Regul."},{"key":"2026090213591904500_cit33","doi-asserted-by":"publisher","first-page":"41","DOI":"10.1016\/0040-4020(61)80097-5","article-title":"The tautomerism of heteroaromatic\ncompounds with five-membered\nrings\u2500I: 5-hydroxyisoxazoles-isoxazol-5-ones","volume":"12","author":"Boulton","year":"1961","journal-title":"Tetrahedron"},{"key":"2026090213591904500_cit34","doi-asserted-by":"publisher","first-page":"1006","DOI":"10.1021\/jm00280a002","article-title":"Utilization\nof operational schemes for analog synthesis in drug design","volume":"15","author":"Topliss","year":"1972","journal-title":"J. Med. Chem."},{"key":"2026090213591904500_cit35","doi-asserted-by":"publisher","first-page":"270","DOI":"10.1007\/s12275-012-2050-4","article-title":"Protein-protein\ninteractions between histidine kinases and response regulators of Mycobacterium tuberculosis H37Rv","volume":"50","author":"Lee","year":"2012","journal-title":"J. Microbiol. (Seoul, Repub. Korea)"},{"key":"2026090213591904500_cit36","doi-asserted-by":"publisher","first-page":"9779","DOI":"10.1073\/pnas.1808874115","article-title":"Arginine-deprivation-induced oxidative\ndamage sterilizes Mycobacterium tuberculosis","volume":"115","author":"Tiwari","year":"2018","journal-title":"Proc. Natl. Acad. Sci. U. S. A."},{"key":"2026090213591904500_cit37","doi-asserted-by":"publisher","first-page":"3258","DOI":"10.1128\/IAI.01449-08","article-title":"Unique roles of\nDosT and DosS in DosR regulon induction and Mycobacterium tuberculosis\ndormancy","volume":"77","author":"Honaker","year":"2009","journal-title":"Infect. Immun."},{"key":"2026090213591904500_cit38","doi-asserted-by":"publisher","first-page":"56","DOI":"10.1111\/mmi.12688","article-title":"Slow growth\nof Mycobacterium tuberculosis\nat acidic pH is regulated by phoPR and host-associated carbon sources","volume":"94","author":"Baker","year":"2014","journal-title":"Mol. Microbiol."},{"key":"2026090213591904500_cit39","doi-asserted-by":"publisher","first-page":"66","DOI":"10.1186\/s12859-016-0923-y","article-title":"SPARTA:\nSimple Program for Automated reference-based bacterial RNA-seq Transcriptome\nAnalysis","volume":"17","author":"Johnson","year":"2016","journal-title":"BMC Bioinf."}],"container-title":["ACS Chemical Biology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acschembio.8b00849","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/acbcct\/article-pdf\/15\/1\/52\/66949630\/acschembio.8b00849.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/acbcct\/article-pdf\/15\/1\/52\/66949630\/acschembio.8b00849.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T17:59:23Z","timestamp":1788371963000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/acbcct\/article\/15\/1\/52\/798165\/Inhibiting-Mycobacterium-tuberculosis-DosRST"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,9,26]]},"references-count":39,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2019,9,26]]},"published-print":{"date-parts":[[2020,1,17]]}},"URL":"https:\/\/doi.org\/10.1021\/acschembio.8b00849","relation":{"has-preprint":[{"id-type":"doi","id":"10.1101\/411793","asserted-by":"object"}]},"ISSN":["1554-8929","1554-8937"],"issn-type":[{"value":"1554-8929","type":"print"},{"value":"1554-8937","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,9,26]]}}}