{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,3,31]],"date-time":"2026-03-31T19:23:38Z","timestamp":1774985018641,"version":"3.50.1"},"posted":{"date-parts":[[2025,1,28]]},"group-title":"Microbiology","reference-count":84,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,1,28]],"date-time":"2025-01-28T00:00:00Z","timestamp":1738022400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,1,28]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Mycobacteria, including\n                  <jats:italic>Mycobacterium tuberculosis<\/jats:italic>\n                  \u2014the etiological agent of tuberculosis\u2014have a unique cell envelope critical for their survival and resistance. The cell envelope\u2019s assembly and maintenance influence permeability, making it a key target against multidrug-resistant strains. Disulfide bond (DSB) formation is crucial for the folding of cell envelope proteins. The DSB pathway in mycobacteria includes two enzymes, DsbA and VKOR, required for survival. Using bioinformatics and cysteine profiling proteomics, we identified cell envelope proteins dependent on DSBs. We validated via\n                  <jats:italic>in vivo<\/jats:italic>\n                  alkylation that key proteins like LamA (MmpS3), PstP, LpqW, and EmbB rely on DSBs for stability. Furthermore, chemical inhibition of VKOR results in phenotypes similar to those of \u0394\n                  <jats:italic>vkor<\/jats:italic>\n                  . Thus, targeting DsbA-VKOR systems could compromise both cell division and mycomembrane integrity. These findings emphasize the potential of DSB inhibition as a novel strategy to combat mycobacterial infections.\n                <\/jats:p>","DOI":"10.1101\/2025.01.27.635063","type":"posted-content","created":{"date-parts":[[2025,1,28]],"date-time":"2025-01-28T23:05:24Z","timestamp":1738105524000},"source":"Crossref","is-referenced-by-count":4,"title":["Disulfide bonds are required for cell division, cell envelope biogenesis and antibiotic resistance proteins in mycobacteria"],"prefix":"10.64898","author":[{"given":"Adrian","family":"Mejia-Santana","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rebecca","family":"Collins","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Emma H.","family":"Doud","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4065-9646","authenticated-orcid":false,"given":"Cristina","family":"Landeta","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2025013102350729000_2025.01.27.635063v1.1","unstructured":"Global tuberculosis report. Global Tuberculosis Report 2021. (World Health Organization, 2021)."},{"key":"2025013102350729000_2025.01.27.635063v1.2","doi-asserted-by":"publisher","DOI":"10.15620\/cdc:82532"},{"key":"2025013102350729000_2025.01.27.635063v1.3","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(21)02724-0\/ATTACHMENT\/4504FC2B-08AE-46D6-BD75-80811DF8B5E6\/MMC1.PDF"},{"key":"2025013102350729000_2025.01.27.635063v1.4","unstructured":"Mirabal A. , B. F. G., N. A. Lung Nontuberculous Mycobacterial Infections. (2024)."},{"key":"2025013102350729000_2025.01.27.635063v1.5","doi-asserted-by":"crossref","unstructured":"Daff\u00e9, M. & Marrakchi, H . Unraveling the Structure of the Mycobacterial Envelope. Microbiol Spectr 7, (2019).","DOI":"10.1128\/microbiolspec.GPP3-0027-2018"},{"key":"2025013102350729000_2025.01.27.635063v1.6","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2020.603382"},{"key":"2025013102350729000_2025.01.27.635063v1.7","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0912952107"},{"key":"2025013102350729000_2025.01.27.635063v1.8","doi-asserted-by":"publisher","DOI":"10.1021\/cb500988r"},{"key":"2025013102350729000_2025.01.27.635063v1.9","doi-asserted-by":"publisher","DOI":"10.1002\/anie.201410341"},{"key":"2025013102350729000_2025.01.27.635063v1.10","doi-asserted-by":"publisher","DOI":"10.1038\/nchembio.1752"},{"key":"2025013102350729000_2025.01.27.635063v1.11","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.73178"},{"key":"2025013102350729000_2025.01.27.635063v1.12","doi-asserted-by":"publisher","DOI":"10.1101\/2023.08.02.551661"},{"key":"2025013102350729000_2025.01.27.635063v1.13","doi-asserted-by":"publisher","DOI":"10.1016\/0092-8674(91)90532-4"},{"key":"2025013102350729000_2025.01.27.635063v1.14","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.90.3.1038"},{"key":"2025013102350729000_2025.01.27.635063v1.15","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.90.15.7084"},{"key":"2025013102350729000_2025.01.27.635063v1.16","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.90.3.1043"},{"key":"2025013102350729000_2025.01.27.635063v1.17","doi-asserted-by":"publisher","DOI":"10.1016\/S0092-8674(00)81016-8"},{"key":"2025013102350729000_2025.01.27.635063v1.18","doi-asserted-by":"publisher","DOI":"10.1016\/j.tibs.2004.04.004"},{"key":"2025013102350729000_2025.01.27.635063v1.19","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0804621105"},{"key":"2025013102350729000_2025.01.27.635063v1.20","doi-asserted-by":"publisher","DOI":"10.1038\/nature02214"},{"key":"2025013102350729000_2025.01.27.635063v1.21","doi-asserted-by":"publisher","DOI":"10.1080\/10409238.2023.2201404"},{"key":"2025013102350729000_2025.01.27.635063v1.22","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbc.2024.107383"},{"key":"2025013102350729000_2025.01.27.635063v1.23","doi-asserted-by":"publisher","DOI":"10.1089\/ars.2010.3558"},{"key":"2025013102350729000_2025.01.27.635063v1.24","doi-asserted-by":"publisher","DOI":"10.1186\/1472-6807-13-23"},{"key":"2025013102350729000_2025.01.27.635063v1.25","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444913017800"},{"key":"2025013102350729000_2025.01.27.635063v1.26","doi-asserted-by":"publisher","DOI":"10.1007\/s13238-013-3033-x"},{"key":"2025013102350729000_2025.01.27.635063v1.27","doi-asserted-by":"publisher","DOI":"10.1038\/emboj.2009.21"},{"key":"2025013102350729000_2025.01.27.635063v1.28","doi-asserted-by":"crossref","unstructured":"Liu, S. et al. Structural basis of antagonizing the vitamin K catalytic cycle for anticoagulation. Science (1979) 371, (2021).","DOI":"10.1126\/science.abc5667"},{"key":"2025013102350729000_2025.01.27.635063v1.29","doi-asserted-by":"publisher","DOI":"10.1128\/jb.00337-22"},{"key":"2025013102350729000_2025.01.27.635063v1.30","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2134250100"},{"key":"2025013102350729000_2025.01.27.635063v1.31","doi-asserted-by":"publisher","DOI":"10.1128\/jb.00137-18"},{"key":"2025013102350729000_2025.01.27.635063v1.32","doi-asserted-by":"crossref","unstructured":"Smith, L. D. & Bertrandt, K. P . Mutations in the TnlO Tet Repressor That Interfere with Induction Location of the Tetracycline-Binding Domain. J. Mol. Biol vol. 203 (1988).","DOI":"10.1016\/0022-2836(88)90120-9"},{"key":"2025013102350729000_2025.01.27.635063v1.33","doi-asserted-by":"publisher","DOI":"10.1128\/mBio"},{"key":"2025013102350729000_2025.01.27.635063v1.34","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2020.02.012"},{"key":"2025013102350729000_2025.01.27.635063v1.35","doi-asserted-by":"publisher","DOI":"10.1038\/nature09472"},{"key":"2025013102350729000_2025.01.27.635063v1.36","doi-asserted-by":"crossref","unstructured":"Dejesus, M. A. et al. Comprehensive essentiality analysis of the Mycobacterium tuberculosis genome via saturating transposon mutagenesis. mBio 8, (2017).","DOI":"10.1128\/mBio.02133-16"},{"key":"2025013102350729000_2025.01.27.635063v1.37","doi-asserted-by":"publisher","DOI":"10.1038\/nature22361"},{"key":"2025013102350729000_2025.01.27.635063v1.38","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M116.754531"},{"key":"2025013102350729000_2025.01.27.635063v1.39","doi-asserted-by":"crossref","unstructured":"Iswahyudi et al. Mycobacterial phosphatase PstP regulates global serine threonine phosphorylation and cell division. Sci Rep 9, (2019).","DOI":"10.1038\/s41598-019-44841-9"},{"key":"2025013102350729000_2025.01.27.635063v1.40","doi-asserted-by":"crossref","unstructured":"Shamma, F. et al. Phosphorylation on PstP regulates cell wall metabolism and antibiotic tolerance in mycobacterium smegmatis. J Bacteriol 203, (2021).","DOI":"10.1128\/JB.00563-20"},{"key":"2025013102350729000_2025.01.27.635063v1.41","doi-asserted-by":"publisher","DOI":"10.1111\/mmi.14951"},{"key":"2025013102350729000_2025.01.27.635063v1.42","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M511709200"},{"key":"2025013102350729000_2025.01.27.635063v1.43","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M112.373415"},{"key":"2025013102350729000_2025.01.27.635063v1.44","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2958.2003.03681.x"},{"key":"2025013102350729000_2025.01.27.635063v1.45","doi-asserted-by":"crossref","unstructured":"Sreevatsan, S. , et al. Ethambutol Resistance in Mycobacterium Tuberculosis: Critical Role of EmbB Mutations. vol. 41 https:\/\/journals.asm.org\/journal\/aac (1997).","DOI":"10.1128\/AAC.41.8.1677"},{"key":"2025013102350729000_2025.01.27.635063v1.46","doi-asserted-by":"publisher","DOI":"10.1002\/1873-3468.12273"},{"key":"2025013102350729000_2025.01.27.635063v1.47","doi-asserted-by":"publisher","DOI":"10.1038\/nmicrobiol.2017.105"},{"key":"2025013102350729000_2025.01.27.635063v1.48","doi-asserted-by":"publisher","DOI":"10.1111\/mmi.14185"},{"key":"2025013102350729000_2025.01.27.635063v1.49","doi-asserted-by":"crossref","unstructured":"Garces, A. et al. EspA acts as a critical mediator of ESX1-dependent virulence in Mycobacterium tuberculosis by affecting bacterial cell wall integrity. PLoS Pathog 6, (2010).","DOI":"10.1371\/journal.ppat.1000957"},{"key":"2025013102350729000_2025.01.27.635063v1.50","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M400811200"},{"key":"2025013102350729000_2025.01.27.635063v1.51","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2006.04.012"},{"key":"2025013102350729000_2025.01.27.635063v1.52","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M110.150094"},{"key":"2025013102350729000_2025.01.27.635063v1.53","doi-asserted-by":"crossref","unstructured":"Alderwick, L. J. et al. The C-terminal domain of the arabinosyltransferase mycobacterium tuberculosis EmbC is a lectin-like carbohydrate binding module. PLoS Pathog 7, (2011).","DOI":"10.1371\/journal.ppat.1001299"},{"key":"2025013102350729000_2025.01.27.635063v1.54","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M113.462036"},{"key":"2025013102350729000_2025.01.27.635063v1.55","doi-asserted-by":"publisher","DOI":"10.1016\/j.jsb.2013.09.022"},{"key":"2025013102350729000_2025.01.27.635063v1.56","doi-asserted-by":"publisher","DOI":"10.1124\/mol.119.118042"},{"key":"2025013102350729000_2025.01.27.635063v1.57","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2020.04.014"},{"key":"2025013102350729000_2025.01.27.635063v1.58","doi-asserted-by":"crossref","unstructured":"Cuthbert, B. J. et al. The structure of Mycobacterium thermoresistibile MmpS5 reveals a conserved disulfide bond across mycobacteria. Metallomics 16, (2024).","DOI":"10.1093\/mtomcs\/mfae011"},{"key":"2025013102350729000_2025.01.27.635063v1.59","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-019-1633-1"},{"key":"2025013102350729000_2025.01.27.635063v1.60","doi-asserted-by":"publisher","DOI":"10.1107\/S1744309105037541"},{"key":"2025013102350729000_2025.01.27.635063v1.61","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.47.9.1309"},{"key":"2025013102350729000_2025.01.27.635063v1.62","doi-asserted-by":"crossref","unstructured":"Tan, Y. Z. et al. Cryo-EM structure of arabinosyltransferase EmbB from Mycobacterium smegmatis. Nat Commun 11, (2020).","DOI":"10.1038\/s41467-020-17202-8"},{"key":"2025013102350729000_2025.01.27.635063v1.63","doi-asserted-by":"publisher","DOI":"10.1126\/science.aba9102"},{"key":"2025013102350729000_2025.01.27.635063v1.64","doi-asserted-by":"publisher","DOI":"10.1016\/j.resmic.2009.07.012"},{"key":"2025013102350729000_2025.01.27.635063v1.65","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2018.12.063"},{"key":"2025013102350729000_2025.01.27.635063v1.66","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.93.23.13048"},{"key":"2025013102350729000_2025.01.27.635063v1.67","doi-asserted-by":"publisher","DOI":"10.1093\/emboj\/18.21.5963"},{"key":"2025013102350729000_2025.01.27.635063v1.68","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M311833200"},{"key":"2025013102350729000_2025.01.27.635063v1.69","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2009.12.060"},{"key":"2025013102350729000_2025.01.27.635063v1.70","doi-asserted-by":"publisher","DOI":"10.1111\/mmi.13565"},{"key":"2025013102350729000_2025.01.27.635063v1.71","doi-asserted-by":"publisher","DOI":"10.1126\/science.1227215"},{"key":"2025013102350729000_2025.01.27.635063v1.72","doi-asserted-by":"publisher","DOI":"10.1016\/j.str.2014.05.008"},{"key":"2025013102350729000_2025.01.27.635063v1.73","doi-asserted-by":"crossref","unstructured":"Iadanza, M. G. et al. Lateral opening in the intact \u03b2-barrel assembly machinery captured by cryo-EM. Nat Commun 7, (2016).","DOI":"10.1038\/ncomms12865"},{"key":"2025013102350729000_2025.01.27.635063v1.74","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.REV120.011201"},{"key":"2025013102350729000_2025.01.27.635063v1.75","doi-asserted-by":"publisher","DOI":"10.1002\/pro.3288"},{"key":"2025013102350729000_2025.01.27.635063v1.76","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.2019"},{"key":"2025013102350729000_2025.01.27.635063v1.77","doi-asserted-by":"crossref","unstructured":"McCracken, N. A. et al. Obtaining Functional Proteomics Insights From Thermal Proteome Profiling Through Optimized Melt Shift Calculation and Statistical Analysis With InflectSSP. Molecular and Cellular Proteomics 22, (2023).","DOI":"10.1016\/j.mcpro.2023.100630"},{"key":"2025013102350729000_2025.01.27.635063v1.78","doi-asserted-by":"crossref","unstructured":"Grecco, G. G. et al. Sex-Dependent Synaptic Remodeling of the Somatosensory Cortex in Mice With Prenatal Methadone Exposure. Advances in Drug and Alcohol Research 2, (2022).","DOI":"10.3389\/adar.2022.10400"},{"key":"2025013102350729000_2025.01.27.635063v1.79","doi-asserted-by":"crossref","unstructured":"Orsburn, B. C . Proteome discoverer-a community enhanced data processing suite for protein informatics. Proteomes 9, (2021).","DOI":"10.3390\/proteomes9010015"},{"key":"2025013102350729000_2025.01.27.635063v1.80","unstructured":"Hallgren, J. et al. DeepTMHMM Predicts Alpha and Beta Transmembrane Proteins Using Deep Neural Networks. https:\/\/dtu.biolib.com\/DeepTMHMM."},{"key":"2025013102350729000_2025.01.27.635063v1.81","doi-asserted-by":"publisher","DOI":"10.1038\/s41587-019-0036-z"},{"key":"2025013102350729000_2025.01.27.635063v1.82","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkp363"},{"key":"2025013102350729000_2025.01.27.635063v1.83","doi-asserted-by":"crossref","unstructured":"Sievers, F. et al. Fast, scalable generation of high-quality protein multiple sequence alignments using Clustal Omega. Mol Syst Biol 7, (2011).","DOI":"10.1038\/msb.2011.75"},{"key":"2025013102350729000_2025.01.27.635063v1.84","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/btt607"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.1101\/2025.01.27.635063","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T07:56:53Z","timestamp":1768463813000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.1101\/2025.01.27.635063"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,1,28]]},"references-count":84,"URL":"https:\/\/doi.org\/10.1101\/2025.01.27.635063","relation":{"is-preprint-of":[{"id-type":"doi","id":"10.1128\/mbio.01083-25","asserted-by":"subject"}]},"subject":[],"published":{"date-parts":[[2025,1,28]]},"subtype":"preprint"}}