{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T11:52:26Z","timestamp":1791460346277,"version":"4.3.4"},"reference-count":45,"publisher":"Oxford University Press (OUP)","license":[{"start":{"date-parts":[[2026,9,23]],"date-time":"2026-09-23T00:00:00Z","timestamp":1790121600000},"content-version":"am","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/pages\/standard-publication-reuse-rights"}],"funder":[{"DOI":"10.13039\/100000060","name":"NIAID","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100012264","name":"NIH","doi-asserted-by":"publisher","award":["R21 AI167027"],"award-info":[{"award-number":["R21 AI167027"]}],"id":[{"id":"10.13039\/501100012264","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100012264","name":"NIH","doi-asserted-by":"publisher","award":["R01AI168088"],"award-info":[{"award-number":["R01AI168088"]}],"id":[{"id":"10.13039\/501100012264","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>Tuberculosis (TB) caused an estimated 10.7 million new cases and 1.23 million deaths in 2024. Antibiotic tolerance, the ability of bacteria to survive bactericidal antibiotics without genetic resistance mutations, contributes to prolonged treatment. Targeting tolerance mechanisms could promote accelerated clearance of Mycobacterium tuberculosis (). Our previous genetic screen identified pstA1, involved in phosphate-specific import.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>PstA1 has previously been implicated in virulence, as mutants lacking this gene exhibit defective survival during phosphate limitation in vitro, in macrophages, and in immunocompetent mice. We evaluated rifampin susceptibility of a pstA1 deletion mutant (\u0394pstA1) using minimum inhibitory concentration (MIC) and time-kill assays, assessed survival during phosphate limitation and in macrophages, and performed differential expression analysis.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>Rifampin MIC was not altered in \u0394pstA1, suggesting that the difference in susceptibility was not due to antibiotic resistance. Time-kill assays revealed a shift in the mean duration of killing (2-log reduction) from 1.6 days in wild type to 1.0 day in \u0394pstA1, and complementation partially restored this phenotype. pstA1 was specifically required for survival in the absence of exogenous inorganic phosphate and was important for growth adaptation in culture without detergent and within macrophages in an interferon-\u03b3-dependent manner. \u0394pstA1 exhibited distinct transcriptional reprogramming with 58 differentially expressed genes, including altered metabolic, DNA damage repair, and secretory pathways.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>These findings identify PstA1 as a contributor to rifampin tolerance and suggest that phosphate transport pathways influence antibiotic susceptibility in .<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1093\/infdis\/jiag483","type":"journal-article","created":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T11:43:37Z","timestamp":1790077417000},"source":"Crossref","is-referenced-by-count":0,"title":["The Phosphate-Specific Transport System Gene\n                    <i>pstA1<\/i>\n                    Contributes to Rifampin Tolerance in\n                    <i>Mycobacterium tuberculosis<\/i>"],"prefix":"10.1093","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2032-3467","authenticated-orcid":false,"given":"Carina","family":"Danchik","sequence":"first","affiliation":[{"name":"Tuberculosis Research Advancement Center, Department of Medicine, Johns Hopkins School of Medicine , Baltimore, Maryland ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-4610-8751","authenticated-orcid":false,"given":"G V Anoushka","family":"Chinmayi","sequence":"additional","affiliation":[{"name":"Tuberculosis Research Advancement Center, Department of Medicine, Johns Hopkins School of Medicine , Baltimore, Maryland ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gi Yong","family":"Lee","sequence":"additional","affiliation":[{"name":"Department of Immunology and Immune Therapeutics, Keck School of Medicine, University of Southern California , Los Angeles, California ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3052-6949","authenticated-orcid":false,"given":"Kathirvel","family":"Maruthai","sequence":"additional","affiliation":[{"name":"Tuberculosis Research Advancement Center, Department of Medicine, Johns Hopkins School of Medicine , Baltimore, Maryland ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6020-4625","authenticated-orcid":false,"given":"Joel S","family":"Bader","sequence":"additional","affiliation":[{"name":"Institute for Computational Medicine, Johns Hopkins University , Baltimore, Maryland ,","place":["USA"]},{"name":"Department of Biomedical Engineering, Johns Hopkins University , Baltimore, Maryland ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hyungjin","family":"Eoh","sequence":"additional","affiliation":[{"name":"Department of Immunology and Immune Therapeutics, Keck School of Medicine, University of Southern California , Los Angeles, California ,","place":["USA"]},{"name":"Department of Ophthalmology, Keck School of Medicine, University of Southern California , Los Angeles, California ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1338-9206","authenticated-orcid":false,"given":"Petros C","family":"Karakousis","sequence":"additional","affiliation":[{"name":"Tuberculosis Research Advancement Center, Department of Medicine, Johns Hopkins School of Medicine , Baltimore, Maryland ,","place":["USA"]},{"name":"Department of International Health, Johns Hopkins Bloomberg School of Public Health , Baltimore, Maryland ,","place":["USA"]},{"name":"Department of Molecular Microbiology and Immunology, Johns Hopkins Bloomberg School of Public Health , Baltimore, Maryland ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2026,9,23]]},"reference":[{"key":"2026100807070079200_jiag483-B1","volume-title":"Global tuberculosis report","year":"2023"},{"key":"2026100807070079200_jiag483-B2","doi-asserted-by":"crossref","first-page":"441","DOI":"10.1038\/s41579-019-0196-3","article-title":"Definitions and guidelines for research on antibiotic persistence","volume":"17","author":"Balaban","year":"2019","journal-title":"Nat Rev Microbiol"},{"key":"2026100807070079200_jiag483-B3","doi-asserted-by":"crossref","first-page":"2721","DOI":"10.1080\/21505594.2021.1990660","article-title":"Insights into the molecular determinants involved in Mycobacterium tuberculosis persistence and their therapeutic implications","volume":"12","author":"Joshi","year":"2021","journal-title":"Virulence"},{"key":"2026100807070079200_jiag483-B4","doi-asserted-by":"crossref","first-page":"320","DOI":"10.1038\/nrmicro.2016.34","article-title":"Distinguishing between resistance, tolerance and persistence to antibiotic treatment","volume":"14","author":"Brauner","year":"2016","journal-title":"Nat Rev Microbiol"},{"key":"2026100807070079200_jiag483-B5","doi-asserted-by":"crossref","first-page":"e0024621","DOI":"10.1128\/Spectrum.00246-21","article-title":"Genetic determinants of intrinsic antibiotic tolerance in Mycobacterium avium","volume":"9","author":"Matern","year":"2021","journal-title":"Microbiol Spectrum"},{"key":"2026100807070079200_jiag483-B6","doi-asserted-by":"crossref","first-page":"1126","DOI":"10.1086\/605700","article-title":"Phosphate depletion: a novel trigger for Mycobacterium tuberculosis persistence","volume":"200","author":"Rifat","year":"2009","journal-title":"J Infect Dis"},{"key":"2026100807070079200_jiag483-B7","doi-asserted-by":"crossref","first-page":"2269","DOI":"10.3390\/microorganisms11092269","article-title":"Functional whole genome screen of nutrient-starved Mycobacterium tuberculosis identifies genes involved in rifampin tolerance","volume":"11","author":"Matern","year":"2023","journal-title":"Microorganisms"},{"key":"2026100807070079200_jiag483-B8","doi-asserted-by":"crossref","first-page":"206","DOI":"10.1016\/0014-5793(96)00953-2","article-title":"Identification of a second Mycobacterium tuberculosis gene cluster encoding proteins of an ABC phosphate transporter","volume":"394","author":"Braibant","year":"1996","journal-title":"FEBS Lett"},{"key":"2026100807070079200_jiag483-B9","doi-asserted-by":"crossref","first-page":"70","DOI":"10.1016\/j.tube.2016.04.010","article-title":"REMap: operon map of M. tuberculosis based on RNA sequence data","volume":"99","author":"Pelly","year":"2016","journal-title":"Tuberculosis"},{"key":"2026100807070079200_jiag483-B10","doi-asserted-by":"crossref","first-page":"e0161467","DOI":"10.1371\/journal.pone.0161467","article-title":"Mycobacterium tuberculosis phosphate uptake system component PstA2 is not required for gene regulation or virulence","volume":"11","author":"Tischler","year":"2016","journal-title":"PLoS One"},{"key":"2026100807070079200_jiag483-B11","doi-asserted-by":"crossref","first-page":"338","DOI":"10.1016\/j.tube.2010.09.004","article-title":"Effect of PstS sub-units or PknD deficiency on the survival of Mycobacterium tuberculosis","volume":"90","author":"Vanzembergh","year":"2010","journal-title":"Tuberculosis (Edinburgh)"},{"key":"2026100807070079200_jiag483-B12","doi-asserted-by":"crossref","first-page":"e0282524","DOI":"10.1128\/mbio.02825-24","article-title":"An exacerbated phosphate starvation response triggers Mycobacterium tuberculosis glycerol utilization at acidic pH","volume":"16","author":"Healy","year":"2025","journal-title":"mBio"},{"key":"2026100807070079200_jiag483-B13","doi-asserted-by":"crossref","first-page":"317","DOI":"10.1128\/IAI.01136-12","article-title":"Mycobacterium tuberculosis requires phosphate-responsive gene regulation to resist host immunity","volume":"81","author":"Tischler","year":"2013","journal-title":"Infect Immunity"},{"key":"2026100807070079200_jiag483-B14","doi-asserted-by":"crossref","first-page":"8327","DOI":"10.1073\/pnas.0503272102","article-title":"Genome-wide requirements for Mycobacterium tuberculosis adaptation and survival in macrophages","volume":"102","author":"Rengarajan","year":"2005","journal-title":"Proc Natl Acad Sci U S A"},{"key":"2026100807070079200_jiag483-B15","doi-asserted-by":"crossref","first-page":"759","DOI":"10.1007\/s00294-016-0604-4","article-title":"Phosphate responsive regulation provides insights for ESX-5 function in Mycobacterium tuberculosis","volume":"62","author":"Elliott","year":"2016","journal-title":"Curr Genet"},{"key":"2026100807070079200_jiag483-B16","doi-asserted-by":"crossref","first-page":"e00778-18","DOI":"10.1128\/mBio.00778-18","article-title":"Mycobacterium tuberculosis pst\/SenX3-RegX3 regulates membrane vesicle production independently of ESX-5 activity","volume":"9","author":"White","year":"2018","journal-title":"mBio"},{"key":"2026100807070079200_jiag483-B17","doi-asserted-by":"crossref","first-page":"265","DOI":"10.1186\/s12866-014-0265-8","article-title":"Karakousis, senX3-independent contribution of regX3 to Mycobacterium tuberculosis virulence","volume":"14","author":"Rifat","year":"2014","journal-title":"BMC Microbiol"},{"key":"2026100807070079200_jiag483-B18","doi-asserted-by":"crossref","first-page":"1125","DOI":"10.1099\/mic.0.077180-0","article-title":"Differential regulation of the two-component regulatory system senX3-regX3 in Mycobacterium tuberculosis","volume":"160","author":"Rifat","year":"2014","journal-title":"Microbiology (Reading)"},{"key":"2026100807070079200_jiag483-B19","doi-asserted-by":"crossref","first-page":"e00494-17","DOI":"10.1128\/mBio.00494-17","article-title":"Mycobacterium tuberculosis PhoY proteins promote persister formation by mediating pst\/SenX3-RegX3 phosphate sensing","volume":"8","author":"Namugenyi","year":"2017","journal-title":"mBio"},{"key":"2026100807070079200_jiag483-B20","doi-asserted-by":"publisher","DOI":"10.1128\/mbio.01467-18","article-title":"ORBIT: a new paradigm for genetic engineering of mycobacterial chromosomes","volume":"9","author":"Murphy","year":"2018","journal-title":"mBio"},{"key":"2026100807070079200_jiag483-B21","doi-asserted-by":"crossref","first-page":"e00039","DOI":"10.1128\/mBio.00039-13","article-title":"The polyphosphate kinase gene ppk2 is required for Mycobacterium tuberculosis inorganic polyphosphate regulation and virulence","volume":"4","author":"Chuang","year":"2013","journal-title":"mBio"},{"key":"2026100807070079200_jiag483-B22","doi-asserted-by":"crossref","first-page":"e02428","DOI":"10.1128\/mBio.02428-14","article-title":"Deficiency of the novel exopolyphosphatase Rv1026\/PPX2 leads to metabolic downshift and altered cell wall permeability in Mycobacterium tuberculosis","volume":"6","author":"Chuang","year":"2015","journal-title":"mBio"},{"key":"2026100807070079200_jiag483-B23","doi-asserted-by":"crossref","first-page":"901","DOI":"10.1016\/S0092-8674(01)00286-0","article-title":"Structural mechanism for rifampicin inhibition of bacterial rna polymerase","volume":"104","author":"Campbell","year":"2001","journal-title":"Cell"},{"key":"2026100807070079200_jiag483-B24","doi-asserted-by":"crossref","first-page":"e0069922","DOI":"10.1128\/msystems.00699-22","article-title":"Mycobacterium tuberculosis requires the outer membrane lipid phthiocerol dimycocerosate for starvation-induced antibiotic tolerance","volume":"8","author":"Block","year":"2023","journal-title":"mSystems"},{"key":"2026100807070079200_jiag483-B25","doi-asserted-by":"crossref","first-page":"5057","DOI":"10.1128\/AAC.04856-14","article-title":"Increased vancomycin susceptibility in mycobacteria: a new approach to identify synergistic activity against multidrug-resistant mycobacteria","volume":"59","author":"Soetaert","year":"2015","journal-title":"Antimicrob Agents Chemother"},{"key":"2026100807070079200_jiag483-B26","doi-asserted-by":"crossref","first-page":"717","DOI":"10.1046\/j.1365-2958.2002.02779.x","article-title":"Evaluation of a nutrient starvation model of Mycobacterium tuberculosis persistence by gene and protein expression profiling","volume":"43","author":"Betts","year":"2002","journal-title":"Mol Microbiol"},{"key":"2026100807070079200_jiag483-B27","doi-asserted-by":"crossref","first-page":"744167","DOI":"10.3389\/fmicb.2021.744167","article-title":"Targeting the Mycobacterium tuberculosis stringent response as a strategy for shortening Tuberculosis treatment","volume":"12","author":"Danchik","year":"2021","journal-title":"Front Microbiol"},{"key":"2026100807070079200_jiag483-B28","doi-asserted-by":"crossref","first-page":"494","DOI":"10.3389\/fmicb.2018.00494","article-title":"Altered Mycobacterium tuberculosis cell wall metabolism and physiology associated with RpoB mutation H526D","volume":"9","author":"Campod\u00f3nico","year":"2018","journal-title":"Front Microbiol"},{"key":"2026100807070079200_jiag483-B29","doi-asserted-by":"crossref","first-page":"eaav2104","DOI":"10.1126\/sciadv.aav2104","article-title":"Inhibiting the stringent response blocks Mycobacterium tuberculosis entry into quiescence and reduces persistence","volume":"5","author":"Dutta","year":"2019","journal-title":"Sci Adv"},{"key":"2026100807070079200_jiag483-B30","doi-asserted-by":"crossref","first-page":"e28076","DOI":"10.1371\/journal.pone.0028076","article-title":"The role of the novel exopolyphosphatase MT0516 in Mycobacterium tuberculosis drug tolerance and persistence","volume":"6","author":"Thayil","year":"2011","journal-title":"PLoS One"},{"key":"2026100807070079200_jiag483-B31","doi-asserted-by":"crossref","first-page":"1607","DOI":"10.1038\/s41564-024-01697-8","article-title":"Propionate prevents loss of the PDIM virulence lipid in Mycobacterium tuberculosis","volume":"9","author":"Mulholland","year":"2024","journal-title":"Nat Microbiol"},{"key":"2026100807070079200_jiag483-B32","doi-asserted-by":"crossref","first-page":"e01934-17","DOI":"10.1128\/JCM.01934-17","article-title":"CLSI methods development and standardization working group best practices for evaluation of antimicrobial susceptibility tests","volume":"56","author":"Humphries","year":"2018","journal-title":"J Clin Microbiol"},{"key":"2026100807070079200_jiag483-B33","doi-asserted-by":"crossref","first-page":"171","DOI":"10.1016\/0378-1119(96)00242-9","article-title":"A Mycobacterium tuberculosis gene cluster encoding proteins of a phosphate transporter homologous to the Escherichia coli Pst system","volume":"176","author":"Braibant","year":"1996","journal-title":"Gene"},{"key":"2026100807070079200_jiag483-B34","doi-asserted-by":"crossref","first-page":"3532","DOI":"10.1099\/mic.0.029199-0","article-title":"Rapid and spontaneous loss of phthiocerol dimycocerosate (PDIM) from Mycobacterium tuberculosis grown in vitro: implications for virulence studies","volume":"155","author":"Domenech","year":"2009","journal-title":"Microbiology (Reading)"},{"key":"2026100807070079200_jiag483-B35","doi-asserted-by":"crossref","first-page":"4778","DOI":"10.1128\/AAC.49.11.4778-4780.2005","article-title":"Differential antibiotic susceptibilities of starved Mycobacterium tuberculosis isolates","volume":"49","author":"Xie","year":"2005","journal-title":"Antimicrob Agents Chemother"},{"key":"2026100807070079200_jiag483-B36","doi-asserted-by":"crossref","first-page":"6460","DOI":"10.1128\/AAC.01139-16","article-title":"Stringent response factors PPX1 and PPK2 play an important role in Mycobacterium tuberculosis metabolism, biofilm formation, and sensitivity to isoniazid in vivo","volume":"60","author":"Chuang","year":"2016","journal-title":"Antimicrob Agents Chemother"},{"key":"2026100807070079200_jiag483-B37","doi-asserted-by":"crossref","first-page":"e2309664121","DOI":"10.1073\/pnas.2309664121","article-title":"Polyphosphate kinase-1 regulates bacterial and host metabolic pathways involved in pathogenesis of Mycobacterium tuberculosis","volume":"121","author":"Chugh","year":"2024","journal-title":"Proc Natl Acad Sci U S A"},{"key":"2026100807070079200_jiag483-B38","doi-asserted-by":"crossref","first-page":"2839","DOI":"10.1128\/JB.00038-13","article-title":"Polyphosphate deficiency in Mycobacterium tuberculosis is associated with enhanced drug susceptibility and impaired growth in Guinea pigs","volume":"195","author":"Singh","year":"2013","journal-title":"J Bacteriol"},{"key":"2026100807070079200_jiag483-B39","doi-asserted-by":"crossref","first-page":"261","DOI":"10.1111\/j.1365-2958.2007.05814.x","article-title":"Polyphosphate kinase is involved in stress-induced mprAB-sigE-rel signalling in mycobacteria","volume":"65","author":"Sureka","year":"2007","journal-title":"Mol Microbiol"},{"key":"2026100807070079200_jiag483-B40","doi-asserted-by":"crossref","first-page":"6554","DOI":"10.1073\/pnas.1219375110","article-title":"Multifunctional essentiality of succinate metabolism in adaptation to hypoxia in Mycobacterium tuberculosis","volume":"110","author":"Eoh","year":"2013","journal-title":"Proc Natl Acad Sci U S A"},{"key":"2026100807070079200_jiag483-B41","doi-asserted-by":"crossref","first-page":"2681","DOI":"10.3389\/fmicb.2017.02681","article-title":"Ultrastructural analysis of cell envelope and accumulation of lipid inclusions in clinical Mycobacterium tuberculosis isolates from sputum, oxidative stress, and iron deficiency","volume":"8","author":"Vijay","year":"2018","journal-title":"Front Microbiol"},{"key":"2026100807070079200_jiag483-B42","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1016\/j.tube.2016.07.002","article-title":"Virulence, biochemistry, morphology and host-interacting properties of detergent-free cultured mycobacteria: an update","volume":"100","author":"Leisching","year":"2016","journal-title":"Tuberculosis"},{"key":"2026100807070079200_jiag483-B43","doi-asserted-by":"crossref","first-page":"461","DOI":"10.1111\/j.1574-6976.2008.00101.x","article-title":"The phosphate regulon and bacterial virulence: a regulatory network connecting phosphate homeostasis and pathogenesis","volume":"32","author":"Lamarche","year":"2008","journal-title":"FEMS Microbiol Rev"},{"key":"2026100807070079200_jiag483-B44","doi-asserted-by":"crossref","first-page":"572","DOI":"10.1016\/j.celrep.2015.12.056","article-title":"Ergothioneine maintains redox and bioenergetic homeostasis essential for drug susceptibility and virulence of Mycobacterium tuberculosis","volume":"14","author":"Saini","year":"2016","journal-title":"Cell Rep"},{"key":"2026100807070079200_jiag483-B45","doi-asserted-by":"crossref","first-page":"11392","DOI":"10.1038\/ncomms11392","article-title":"Thiol reductive stress induces cellulose-anchored biofilm formation in Mycobacterium tuberculosis","volume":"7","author":"Trivedi","year":"2016","journal-title":"Nat Commun"}],"container-title":["The Journal of Infectious Diseases"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/academic.oup.com\/jid\/advance-article-pdf\/doi\/10.1093\/infdis\/jiag483\/71334513\/jiag483.pdf","content-type":"application\/pdf","content-version":"am","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/jid\/advance-article-pdf\/doi\/10.1093\/infdis\/jiag483\/71334513\/jiag483.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T11:07:12Z","timestamp":1791457632000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/jid\/advance-article\/doi\/10.1093\/infdis\/jiag483\/8831301"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,23]]},"references-count":45,"URL":"https:\/\/doi.org\/10.1093\/infdis\/jiag483","relation":{},"ISSN":["0022-1899","1537-6613"],"issn-type":[{"value":"0022-1899","type":"print"},{"value":"1537-6613","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,23]]},"article-number":"jiag483"}}