{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,14]],"date-time":"2026-08-14T15:02:47Z","timestamp":1786719767003,"version":"build-2736575974"},"reference-count":52,"publisher":"American Chemical Society (ACS)","issue":"8","license":[{"start":{"date-parts":[[2026,8,5]],"date-time":"2026-08-05T00:00:00Z","timestamp":1785888000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100018833","name":"Agence de l'innovation de D\u00e9fense","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100018833","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,8,14]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Trimethoprim\u2013sulfamethoxazole (SXT) remains central to melioidosis eradication therapy, yet the genetic basis of resistance evolution in Burkholderia pseudomallei is not fully defined. Using Burkholderia thailandensis as a biosafe surrogate, we subjected populations to stepwise in vitro evolution under increasing SXT concentrations. Whole-genome sequencing revealed rapid population diversification followed by selective sweeps leading to fixation of fitter resistant variants. Early adaptation involved mutations in regulators of RND efflux systems and folate-associated genes, consistent with multifactorial resistance. Despite this heterogeneity, all evolutionary trajectories converged on a single nonsynonymous substitution, I99L, in dihydrofolate reductase (DHFR\/FolA), which rose to fixation at high SXT concentrations. This substitution mirrors changes reported in SXT-resistant B. pseudomallei isolates from chronic infections. Allelic reconstruction demonstrated that Bt-DHFR(I99L) is sufficient to confer a 4- to 16-fold increase in trimethoprim MIC. Biochemical and structural analyses showed preserved catalytic activity but reduced trimethoprim binding, consistent with an allosteric resistance mechanism. Together, these findings establish DHFR as a dominant evolutionary target under SXT pressure and support B. thailandensis as a robust model for dissecting resistance evolution in B. pseudomallei.<\/jats:p>","DOI":"10.1021\/acsinfecdis.5c01132","type":"journal-article","created":{"date-parts":[[2026,8,6]],"date-time":"2026-08-06T01:02:15Z","timestamp":1785978135000},"page":"2627-2641","source":"Crossref","is-referenced-by-count":0,"title":["Evolutionary\nConvergence on a Dihydrofolate Reductase\nMutation Drives Trimethoprim\u2013Sulfamethoxazole Resistance in\n                    <i>Burkholderia thailandensis<\/i>"],"prefix":"10.1021","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2106-7901","authenticated-orcid":true,"given":"Sophie","family":"Guillier","sequence":"first","affiliation":[{"name":"Institut de Recherche Biome\u0301dicale des Arme\u0301es (IRBA) , , ,","place":["Bre\u0301tigny-Sur-Orge, France, 91220"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christine","family":"Lazennec-Schurdevin","sequence":"additional","affiliation":[{"name":"CNRS, Ecole Polytechnique, Institut Polytechnique de Paris , , ,","place":["Palaiseau, France, 91120"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lou","family":"Mondange","sequence":"additional","affiliation":[{"name":"Institut de Recherche Biome\u0301dicale des Arme\u0301es (IRBA) , , ,","place":["Bre\u0301tigny-Sur-Orge, France, 91220"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9067-7186","authenticated-orcid":true,"given":"Ve\u0301ronique","family":"Sarilar","sequence":"additional","affiliation":[{"name":"Institut de Recherche Biome\u0301dicale des Arme\u0301es (IRBA) , ,","place":["Bre\u0301tigny-Sur-Orge, France, 91220"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Me\u0301lissa","family":"Marchandeau","sequence":"additional","affiliation":[{"name":"Pharmacie Centrale des Arme\u0301es , ,","place":["Fleury-les-Aubrais, France, 45404"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chloe\u0301","family":"Lemoigne","sequence":"additional","affiliation":[{"name":"Institut de Recherche Biome\u0301dicale des Arme\u0301es (IRBA) , , ,","place":["Bre\u0301tigny-Sur-Orge, France, 91220"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ombeline","family":"Lamer","sequence":"additional","affiliation":[{"name":"Institut de Recherche Biome\u0301dicale des Arme\u0301es (IRBA) , , ,","place":["Bre\u0301tigny-Sur-Orge, France, 91220"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mathilde","family":"Lescat","sequence":"additional","affiliation":[{"name":"Institut de Recherche Biome\u0301dicale des Arme\u0301es (IRBA) , , ,","place":["Bre\u0301tigny-Sur-Orge, France, 91220"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emmanuelle","family":"Schmitt","sequence":"additional","affiliation":[{"name":"CNRS, Ecole Polytechnique, Institut Polytechnique de Paris , , ,","place":["Palaiseau, France, 91120"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yves","family":"Mechulam","sequence":"additional","affiliation":[{"name":"CNRS, Ecole Polytechnique, Institut Polytechnique de Paris , , ,","place":["Palaiseau, France, 91120"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Julian","family":"Garrec","sequence":"additional","affiliation":[{"name":"UCP, ENSTA, Institut Polytechnique de Paris , ,","place":["Palaiseau, France, 91120"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fabrice","family":"Biot","sequence":"additional","affiliation":[{"name":"Direction Centrale du Service de Sante\u0301 des Arme\u0301es (DCSSA) , ,","place":["Paris, France, 75015"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,8,5]]},"reference":[{"key":"2026081410070911700_cit1","doi-asserted-by":"publisher","first-page":"155","DOI":"10.1038\/s41579-023-00972-5","article-title":"Burkholderia Pseudomallei and Melioidosis","volume":"22","author":"Meumann","year":"2024","journal-title":"Nat. Rev. Microbiol"},{"key":"2026081410070911700_cit2","doi-asserted-by":"publisher","first-page":"29","DOI":"10.1016\/j.ijid.2016.02.103","article-title":"Predicted\nDistribution of B. Pseudomallei and Burden of Melioidosis in South\nAsia and Worldwide","volume":"45","author":"Limmathurotsakul","year":"2016","journal-title":"International Journal of\nInfectious Diseases"},{"issue":"11","key":"2026081410070911700_cit3","doi-asserted-by":"publisher","first-page":"926","DOI":"10.3390\/pathogens13110926","article-title":"The Evolving Global Epidemiology\nof Human Melioidosis:\nA Narrative Review","volume":"13","author":"Norman","year":"2024","journal-title":"Pathogens"},{"key":"2026081410070911700_cit4","doi-asserted-by":"publisher","first-page":"650","DOI":"10.1128\/JCM.02239-13","article-title":"Recurrent\nMelioidosis in the Darwin\nProspective Melioidosis Study: Improving Therapies Mean that Relapse\nCases Are Now Rare","volume":"52","author":"Sarovich","year":"2014","journal-title":"J. Clin Microbiol"},{"issue":"4","key":"2026081410070911700_cit5","doi-asserted-by":"publisher","first-page":"310","DOI":"10.1016\/j.ijantimicag.2014.01.005","article-title":"Treatment\nand Prophylaxis of Melioidosis","volume":"43","author":"Dance","year":"2014","journal-title":"Int. J. Antimicrob.\nAgents"},{"issue":"9","key":"2026081410070911700_cit6","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0008659","article-title":"2020 Review\nand Revision of the 2015 Darwin Melioidosis Treatment Guideline; Paradigm\nDrift Not Shift","volume":"14","author":"Sullivan","year":"2020","journal-title":"PLoS Negl Trop Dis"},{"issue":"1","key":"2026081410070911700_cit7","doi-asserted-by":"publisher","first-page":"74","DOI":"10.1186\/s12941-023-00620-z","article-title":"Efficacy\nand Safety of Co-Trimoxazole\nin Eradication Phase of Melioidosis; Systematic Review","volume":"22","author":"Keragala","year":"2023","journal-title":"Ann. Clin Microbiol Antimicrob"},{"issue":"5","key":"2026081410070911700_cit8","doi-asserted-by":"publisher","first-page":"557","DOI":"10.1016\/j.ijantimicag.2015.01.006","article-title":"Trimethoprim\/Sulfamethoxazole\nResistance\nin Clinical Isolates of Burkholderia Pseudomallei from Thailand","volume":"45","author":"Saiprom","year":"2015","journal-title":"Int. J. Antimicrob. Agents"},{"issue":"4","key":"2026081410070911700_cit9","doi-asserted-by":"publisher","first-page":"368","DOI":"10.1016\/j.ijantimicag.2014.06.003","article-title":"Trimethoprim\/Sulfamethoxazole Resistance\nin Burkholderia Pseudomallei","volume":"44","author":"Dance","year":"2014","journal-title":"Int. J. Antimicrob. Agents"},{"issue":"5","key":"2026081410070911700_cit10","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.01357-17","article-title":"Mechanisms\nof Resistance to Folate Pathway Inhibitors in Burkholderia Pseudomallei: Deviation from the Norm","volume":"8","author":"Podnecky","year":"2017","journal-title":"mBio"},{"issue":"2","key":"2026081410070911700_cit11","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0008913","article-title":"Genomic and\nRT-qPCR Analysis of Trimethoprim-Sulfamethoxazole and Meropenem Resistance\nin Burkholderia Pseudomallei Clinical Isolates","volume":"15","author":"Schnetterle","year":"2021","journal-title":"PLoS Negl Trop Dis"},{"issue":"6","key":"2026081410070911700_cit12","doi-asserted-by":"publisher","first-page":"1029","DOI":"10.1093\/jac\/dki151","article-title":"Trimethoprim\/Sulfamethoxazole\nResistance in Clinical Isolates of Burkholderia Pseudomallei","volume":"55","author":"Wuthiekanun","year":"2005","journal-title":"J. Antimicrob. Chemother."},{"key":"2026081410070911700_cit13","doi-asserted-by":"publisher","first-page":"2","DOI":"10.1128\/mBio.00356-17","article-title":"Within-Host\nEvolution of Burkholderia Pseudomallei during Chronic\nInfection of Seven\nAustralasian Cystic Fibrosis Patients","volume":"8","author":"Viberg","year":"2017","journal-title":"mBio"},{"issue":"12","key":"2026081410070911700_cit14","doi-asserted-by":"publisher","first-page":"1389","DOI":"10.2217\/fmb.12.116","article-title":"Mechanisms\nof Antibiotic Resistance in Burkholderia Pseudomallei: Implications for Treatment of Melioidosis","volume":"7","author":"Schweizer","year":"2012","journal-title":"Future Microbiology"},{"issue":"46","key":"2026081410070911700_cit15","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2406771121","article-title":"Combatting\nMelioidosis with Chemical Synthetic Lethality","volume":"121","author":"Zhang","year":"2024","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"issue":"5","key":"2026081410070911700_cit16","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0036507","article-title":"Evolution of Burkholderia Pseudomallei in Recurrent Melioidosis.","volume":"7","author":"Hayden","year":"2012","journal-title":"PLoS One"},{"issue":"6","key":"2026081410070911700_cit17","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.00010-17","article-title":"Antibiotic Resistance\nMarkers in Burkholderia Pseudomallei Strain Bp1651 Identified by Genome Sequence\nAnalysis","volume":"61","author":"Bugrysheva","year":"2017","journal-title":"Antimicrob. Agents Chemother."},{"issue":"9","key":"2026081410070911700_cit18","doi-asserted-by":"publisher","first-page":"1156","DOI":"10.1099\/mic.0.000691","article-title":"Molecular\nDeterminants of Burkholderia Pseudomallei BpeEF-OprC\nEfflux Pump Expression","volume":"164","author":"Rhodes","year":"2018","journal-title":"Microbiology\n(Reading)"},{"issue":"6","key":"2026081410070911700_cit19","doi-asserted-by":"publisher","first-page":"429","DOI":"10.1177\/108705710100600609","article-title":"High-Density Miniaturized Thermal\nShift Assays as a General Strategy for Drug Discovery","volume":"6","author":"Pantoliano","year":"2001","journal-title":"J. Biomol Screen"},{"issue":"3","key":"2026081410070911700_cit20","doi-asserted-by":"publisher","first-page":"706","DOI":"10.1002\/prot.22383","article-title":"Structural\nComparison of Chromosomal and Exogenous Dihydrofolate Reductase from\nStaphylococcus Aureus in Complex with the Potent Inhibitor Trimethoprim","volume":"76","author":"Heaslet","year":"2009","journal-title":"Proteins: Struct., Funct., Bioinf."},{"issue":"1","key":"2026081410070911700_cit21","doi-asserted-by":"publisher","first-page":"119","DOI":"10.1146\/annurev.biophys.33.110502.133613","article-title":"Structure,\nDynamics, and Catalytic\nFunction of Dihydrofolate Reductase","volume":"33","author":"Schnell","year":"2004","journal-title":"Annu. Rev.\nBiophys. Biomol. Struct."},{"issue":"24","key":"2026081410070911700_cit22","doi-asserted-by":"publisher","first-page":"6715","DOI":"10.1021\/acs.jcim.2c00507","article-title":"DHFR Mutants\nModulate Their Synchronized\nDynamics with the Substrate by Shifting Hydrogen Bond Occupancies","volume":"62","author":"Cetin","year":"2022","journal-title":"J. Chem. Inf. Model."},{"issue":"1","key":"2026081410070911700_cit23","doi-asserted-by":"publisher","first-page":"2949","DOI":"10.1038\/s41467-021-23191-z","article-title":"A Trimethoprim Derivative\nImpedes Antibiotic Resistance Evolution","volume":"12","author":"Manna","year":"2021","journal-title":"Nat.\nCommun."},{"issue":"8","key":"2026081410070911700_cit24","doi-asserted-by":"publisher","first-page":"2710","DOI":"10.1021\/bi00008a039","article-title":"Isomorphous Crystal\nStructures of Escherichia Coli Dihydrofolate Reductase Complexed with\nFolate, 5-Deazafolate, and 5,10-Dideazatetrahydrofolate: Mechanistic\nImplications","volume":"34","author":"Reyes","year":"1995","journal-title":"Biochemistry"},{"issue":"13","key":"2026081410070911700_cit25","doi-asserted-by":"publisher","first-page":"4085","DOI":"10.1021\/bi00387a052","article-title":"Construction\nand Evaluation of the\nKinetic Scheme Associated with Dihydrofolate Reductase from Escherichia Coli","volume":"26","author":"Fierke","year":"1987","journal-title":"Biochemistry"},{"issue":"22","key":"2026081410070911700_cit26","doi-asserted-by":"publisher","first-page":"13650","DOI":"10.1016\/S0021-9258(18)33497-5","article-title":"Crystal Structures\nof Escherichia Coli and Lactobacillus Casei Dihydrofolate Reductase Refined at 1.7 A Resolution. I. General\nFeatures and Binding of Methotrexate","volume":"257","author":"Bolin","year":"1982","journal-title":"J. Biol.\nChem."},{"issue":"3","key":"2026081410070911700_cit27","doi-asserted-by":"publisher","first-page":"586","DOI":"10.1021\/bi962337c","article-title":"Loop and Subdomain Movements in the\nMechanism of Escherichia Coli Dihydrofolate Reductase: Crystallographic Evidence,","volume":"36","author":"Sawaya","year":"1997","journal-title":"Biochemistry"},{"issue":"7","key":"2026081410070911700_cit28","doi-asserted-by":"publisher","first-page":"1533","DOI":"10.1093\/molbev\/msz086","article-title":"High-Order Epistasis\nin Catalytic Power of Dihydrofolate Reductase Gives Rise to a Rugged\nFitness Landscape in the Presence of Trimethoprim Selection","volume":"36","author":"Tamer","year":"2019","journal-title":"Mol. Biol. Evol."},{"issue":"4","key":"2026081410070911700_cit29","doi-asserted-by":"publisher","first-page":"1128","DOI":"10.1128\/AAC.48.4.1128-1135.2004","article-title":"BpeAB-OprB, a Multidrug\nEfflux Pump in Burkholderia Pseudomallei","volume":"48","author":"Chan","year":"2004","journal-title":"Antimicrob. Agents Chemother."},{"issue":"2","key":"2026081410070911700_cit30","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0016892","article-title":"Involvement\nof the Efflux Pumps in Chloramphenicol Selected Strains of Burkholderia Thailandensis: Proteomic and Mechanistic Evidence","volume":"6","author":"Biot","year":"2011","journal-title":"PLoS One"},{"issue":"4","key":"2026081410070911700_cit31","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.00500-17","article-title":"Metabolic Compensation of Fitness\nCosts Is a General Outcome for Antibiotic-Resistant Pseudomonas Aeruginosa Mutants Overexpressing Efflux Pumps","volume":"8","author":"Olivares Pacheco","year":"2017","journal-title":"mBio"},{"issue":"7","key":"2026081410070911700_cit32","doi-asserted-by":"publisher","first-page":"633","DOI":"10.1517\/17460441.2012.688949","article-title":"Understanding\nEfflux in Gram-Negative Bacteria: Opportunities for Drug Discovery","volume":"7","author":"Schweizer","year":"2012","journal-title":"Expert Opinion on Drug Discovery"},{"issue":"1","key":"2026081410070911700_cit33","doi-asserted-by":"publisher","first-page":"1003","DOI":"10.1038\/s41467-018-03447-x","article-title":"Mutual Potentiation\nDrives Synergy\nbetween Trimethoprim and Sulfamethoxazole","volume":"9","author":"Minato","year":"2018","journal-title":"Nat.\nCommun."},{"issue":"6","key":"2026081410070911700_cit34","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.01769-17","article-title":"Subversion of Metabolic Wasting as\nthe Mechanism for folM -Linked Sulfamethoxazole Resistance","volume":"8","author":"Minato","year":"2017","journal-title":"mBio"},{"issue":"15","key":"2026081410070911700_cit35","doi-asserted-by":"publisher","first-page":"4839","DOI":"10.1021\/acs.jcim.3c00818","article-title":"Kinetic Barrier to\nEnzyme Inhibition Is Manipulated by Dynamical Local Interactions in E. Coli DHFR","volume":"63","author":"Cetin","year":"2023","journal-title":"J. Chem. Inf. Model."},{"issue":"9","key":"2026081410070911700_cit36","doi-asserted-by":"publisher","first-page":"2387","DOI":"10.1093\/molbev\/msu191","article-title":"Strength of Selection Pressure Is an Important Parameter\nContributing to the Complexity of Antibiotic Resistance Evolution","volume":"31","author":"Oz","year":"2014","journal-title":"Mol. Biol. Evol."},{"key":"2026081410070911700_cit37","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.64518","article-title":"Development\nof Antibacterial Compounds That Constrain Evolutionary Pathways to\nResistance","volume":"10","author":"Zhang","year":"2021","journal-title":"eLife"},{"key":"2026081410070911700_cit38","doi-asserted-by":"publisher","DOI":"10.1101\/2025.01.23.634126","article-title":"Exploring Antibiotic\nResistance in Diverse Homologs of the Dihydrofolate Reductase Protein\nFamily through Broad Mutational Scanning","author":"Romanowicz","year":"2025","journal-title":"Synth.\nBiol."},{"key":"2026081410070911700_cit39","doi-asserted-by":"publisher","DOI":"10.1016\/j.insi.2025.100132","article-title":"Harnessing AI-Powered Computational\nDrug Design to\nCombat Antimicrobial Resistance: A Comprehensive Review of Challenges,\nOpportunities, and Future Directions","volume":"1","author":"Santhiya","year":"2025","journal-title":"In Silico\nResearch in Biomedicine"},{"issue":"21","key":"2026081410070911700_cit40","doi-asserted-by":"publisher","first-page":"5962","DOI":"10.1016\/j.cell.2025.07.033","article-title":"A Generative Deep Learning Approach to de Novo Antibiotic\nDesign","volume":"188","author":"Krishnan","year":"2025","journal-title":"Cell"},{"key":"2026081410070911700_cit41","doi-asserted-by":"publisher","first-page":"165","DOI":"10.1007\/978-1-4939-0554-6_12","article-title":"Identification of Mutations in Laboratory-Evolved\nMicrobes from Next-Generation\nSequencing Data Using Breseq","volume-title":"Engineering\nand Analyzing Multicellular Systems","author":"Deatherage","year":"2014"},{"issue":"8","key":"2026081410070911700_cit42","doi-asserted-by":"publisher","DOI":"10.1128\/spectrum.00748-24","article-title":"Unraveling the Role\nof Toxin-Antitoxin Systems in Burkholderia Pseudomallei: Exploring Bacterial Pathogenesis\nand Interactions within the HigBA Families","volume":"12","author":"Chapartegui-Gonza\u0301lez","year":"2024","journal-title":"Microbiol. Spectr"},{"key":"2026081410070911700_cit43","doi-asserted-by":"publisher","first-page":"1001","DOI":"10.1016\/j.profoo.2011.09.150","article-title":"Quantifying the\nCombined Effect of Salt and Temperature\non the Growth of Listeria Strains Isolated from Salmon\nand Salmon Processing Environments","volume":"1","author":"Sk\u00e5ra","year":"2011","journal-title":"Procedia\nFood Science"},{"issue":"47","key":"2026081410070911700_cit44","doi-asserted-by":"publisher","first-page":"15594","DOI":"10.1021\/bi051514j","article-title":"Initiator tRNA Binding\nby e\/aIF5B, the Eukaryotic\/Archaeal Homologue of Bacterial Initiation\nFactor IF2","volume":"44","author":"Guillon","year":"2005","journal-title":"Biochemistry"},{"issue":"16","key":"2026081410070911700_cit45","doi-asserted-by":"publisher","first-page":"3757","DOI":"10.1021\/bi00259a006","article-title":"Kinetic Mechanism\nof the Reaction Catalyzed by Dihydrofolate\nReductase from Escherichia Coli","volume":"21","author":"Stone","year":"1982","journal-title":"Biochemistry"},{"key":"2026081410070911700_cit46","unstructured":"PomBase.\n           - Reference - PMID:37507029 -  Binding of human Cdc123 to\neIF2\u03b3. https:\/\/pombase.org\/reference\/PMID:37507029 (accessed 2025\u201309\u201312)."},{"issue":"2","key":"2026081410070911700_cit47","doi-asserted-by":"publisher","first-page":"125","DOI":"10.1107\/S0907444909047337","article-title":"XDS","volume":"66","author":"Kabsch","year":"2010","journal-title":"Acta Crystallogr. D Biol. Crystallogr."},{"issue":"Pt 6","key":"2026081410070911700_cit48","doi-asserted-by":"publisher","first-page":"449","DOI":"10.1107\/S2059798323003595","article-title":"The CCP4 Suite:\nIntegrative Software for Macromolecular Crystallography","volume":"79","author":"Agirre","year":"2023","journal-title":"Acta Crystallogr. D Struct Biol."},{"issue":"Pt 4","key":"2026081410070911700_cit49","doi-asserted-by":"publisher","first-page":"352","DOI":"10.1107\/S0907444912001308","article-title":"Towards\nAutomated Crystallographic\nStructure Refinement with Phenix.Refine","volume":"68","author":"Afonine","year":"2012","journal-title":"Acta\nCrystallogr. D Biol. Crystallogr."},{"issue":"Pt 4","key":"2026081410070911700_cit50","doi-asserted-by":"publisher","first-page":"486","DOI":"10.1107\/S0907444910007493","article-title":"Features and\nDevelopment\nof Coot","volume":"66","author":"Emsley","year":"2010","journal-title":"Acta Crystallogr. D Biol. Crystallogr."},{"issue":"11","key":"2026081410070911700_cit51","doi-asserted-by":"publisher","DOI":"10.1002\/pro.4792","article-title":"UCSF ChimeraX:\nTools for Structure\nBuilding and Analysis","volume":"32","author":"Meng","year":"2023","journal-title":"Protein Sci."},{"issue":"13","key":"2026081410070911700_cit52","doi-asserted-by":"publisher","first-page":"3320","DOI":"10.1093\/nar\/gkg556","article-title":"ESPript\/ENDscript:\nExtracting and Rendering Sequence and 3D Information from Atomic Structures\nof Proteins","volume":"31","author":"Gouet","year":"2003","journal-title":"Nucleic Acids Res."}],"container-title":["ACS Infectious Diseases"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acsinfecdis.5c01132","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/aidcbc\/article-pdf\/12\/8\/2627\/66422485\/acsinfecdis.5c01132.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/aidcbc\/article-pdf\/12\/8\/2627\/66422485\/acsinfecdis.5c01132.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,14]],"date-time":"2026-08-14T14:20:02Z","timestamp":1786717202000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/aidcbc\/article\/12\/8\/2627\/5246586\/Evolutionary-Convergence-on-a-Dihydrofolate"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,5]]},"references-count":52,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2026,8,5]]},"published-print":{"date-parts":[[2026,8,14]]}},"URL":"https:\/\/doi.org\/10.1021\/acsinfecdis.5c01132","relation":{},"ISSN":["2373-8227"],"issn-type":[{"value":"2373-8227","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,5]]}}}