{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,28]],"date-time":"2026-02-28T04:25:54Z","timestamp":1772252754612,"version":"3.50.1"},"reference-count":80,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2022,3,18]],"date-time":"2022-03-18T00:00:00Z","timestamp":1647561600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>The intracellular pathogen Shigella flexneri, which is the causative agent of bacillary dysentery, significantly influences the worldwide implication of diarrheal infections, consequentially causing about 1.1 million deaths each year. Due to a nonavailability of an authorized vaccine and the upsurge of multidrug resistance amongst Shigella strains, there has been a huge demand for further genetic analyses which could help in the advancement of new\/improved drugs, and finding vaccine candidates against the pathogen. The present study aims to illustrate the role of the yfiB gene in Shigella virulence, part of the periplasmic YfiBNR tripartite signalling system. This system is involved in the regulation of cyclic-di-GMP levels inside the bacterial cells, a vital messenger molecule impacting varied cellular processes such as biofilm formation, cytotoxicity, motility, synthesis of exopolysaccharide, and other virulence mechanisms such as adhesion and invasion of the bacteria. Through a combination of genetic, biochemical, and virulence assays, we show how knocking out the yfiB gene can disrupt the entire YfiBNR system and affect the native c-di-GMP levels. We found that this subsequently causes a negative effect on the biofilm formation, bacterial invasion, host\u2013surface attachment, and the overall virulence of Shigella. This study also carried out a structural and functional assessment of the YfiB protein and determined critical amino acid residues, essential for proper functioning of this signalling system. The present work improves our understanding of the in vivo persistence and survival of Shigella, brings light to the c-di-GMP led regulation of Shigella virulence, and provides a prospective new target to design anti-infection drugs and vaccines against S. flexneri and other bacterial pathogens.<\/jats:p>","DOI":"10.3390\/microorganisms10030653","type":"journal-article","created":{"date-parts":[[2022,3,20]],"date-time":"2022-03-20T21:34:27Z","timestamp":1647812067000},"page":"653","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["YfiB: An Outer Membrane Protein Involved in the Virulence of Shigella flexneri"],"prefix":"10.3390","volume":"10","author":[{"given":"Tanuka","family":"Sen","sequence":"first","affiliation":[{"name":"Division of Biomedical Science and Biochemistry, Research School of Biology, The Australian National University, Canberra, ACT 2601, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Naresh K.","family":"Verma","sequence":"additional","affiliation":[{"name":"Division of Biomedical Science and Biochemistry, Research School of Biology, The Australian National University, Canberra, ACT 2601, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,3,18]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"801","DOI":"10.1016\/S0140-6736(17)33296-8","article-title":"Shigellosis","volume":"391","author":"Kotloff","year":"2018","journal-title":"Lancet"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1016\/j.femsre.2003.07.002","article-title":"Shigella flexneri infection: Pathogenesis and vaccine development","volume":"28","author":"Jennison","year":"2004","journal-title":"FEMS Microbiol. Rev."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"909","DOI":"10.1016\/S1473-3099(17)30276-1","article-title":"Estimates of global, regional, and national morbidity, mortality, and aetiologies of diarrhoeal diseases: A systematic analysis for the Global Burden of Disease Study","volume":"17","author":"Troeger","year":"2017","journal-title":"Lancet Infect. Dis."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"449","DOI":"10.1097\/QCO.0000000000000475","article-title":"Recent insights into Shigella: A major contributor to the global diarrhoeal disease burden","volume":"31","author":"Baker","year":"2018","journal-title":"Curr. Opin. Infect. Dis."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Von Seidlein, L., Kim, D.R., Ali, M., Lee, H., Wang, X., Thiem, V.D., Canh, D.G., Chaicumpa, W., Agtini, M.D., and Hossain, A. (2006). A Multicentre Study of Shigella Diarrhoea in Six Asian Countries: Disease Burden, Clinical Manifestations, and Microbiology. PLoS Med., 3.","DOI":"10.1371\/journal.pmed.0030353"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"275","DOI":"10.1007\/s12223-020-00773-w","article-title":"Shigella flexneri: An emerging pathogen","volume":"65","author":"Nisa","year":"2020","journal-title":"Folia Microbiol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1186\/s13073-020-00824-4","article-title":"The identification of novel immunogenic antigens as potential Shigella vaccine components","volume":"13","author":"Liang","year":"2021","journal-title":"Genome Med."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"7372","DOI":"10.1016\/j.vaccine.2017.09.083","article-title":"Considerations for using ETEC and Shigella disease burden estimates to guide vaccine development strategy","volume":"37","author":"Hosangadi","year":"2019","journal-title":"Vaccine"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"3137","DOI":"10.2147\/IDR.S219755","article-title":"Shigella: Antibiotic-resistance mechanisms and new horizons for treatment","volume":"12","author":"Ranjbar","year":"2019","journal-title":"Infect. Drug Resist."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1128\/microbiolspec.BAI-0023-2019","article-title":"Shigella Pathogenesis: New Insights through Advanced Methodologies","volume":"7","author":"Schnupf","year":"2019","journal-title":"Microbiol. Spectr."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"134","DOI":"10.1128\/CMR.00032-07","article-title":"Molecular Pathogenesis of Shigella spp.: Controlling Host Cell Signaling, Invasion, and Death by Type III Secretion","volume":"21","author":"Schroeder","year":"2008","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"267","DOI":"10.1111\/j.1574-6968.1999.tb08854.x","article-title":"Adhesion and invasion of a mutant Shigella flexneri to an eukaryotic cell line in absence of the 220-kb virulence plasmid","volume":"181","author":"Guhathakurta","year":"1999","journal-title":"FEMS Microbiol. Lett."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"680","DOI":"10.1111\/j.1365-2958.2006.05547.x","article-title":"Shigella chromosomal IpaH proteins are secreted via the type III secretion system and act as effectors","volume":"63","author":"Ashida","year":"2007","journal-title":"Mol. Microbiol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"675","DOI":"10.1046\/j.1365-2958.1999.01627.x","article-title":"A system for identifying post-invasion functions of invasion genes: Requirements for the Mxi\u2013Spa type III secretion pathway of Shigella flexneri in intercellular dissemination","volume":"34","author":"Schuch","year":"1999","journal-title":"Mol. Microbiol."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Reeves, A.Z., Costa, S.C., and Lesser, C.F. (2016). Molecular Approaches to Identifying Type 3 Secreted Effectors: A Shigella Case Study. Shigella, Caister Academic Press.","DOI":"10.21775\/9781910190197.13"},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Qin, J., Doyle, M.T., Tran, E.N.H., and Morona, R. (2020). The virulence domain of Shigella IcsA contains a subregion with specific host cell adhesion function. PLoS ONE, 15.","DOI":"10.1371\/journal.pone.0227425"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Zhu, Z., Wang, W., Cao, M., Zhu, Q., Ma, T., Zhang, Y., Liu, G., Zhou, Z., Li, B., and Shi, Y. (2021). Virulence Factors and Molecular Characteristics of Shigella Flexneri Isolated from Calves with Diarrhea. BMC Microbiol., 21.","DOI":"10.1186\/s12866-021-02277-0"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"e00751-19","DOI":"10.1128\/mSphere.00751-19","article-title":"Shigella flexneri adherence factor expression in in vivo-like conditions","volume":"4","author":"Chanin","year":"2019","journal-title":"mSphere"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"e01067-16","DOI":"10.1128\/IAI.01067-16","article-title":"Analysis of Shigella flexneri resistance, biofilm formation, and transcriptional profile in response to bile salts","volume":"85","author":"Nickerson","year":"2017","journal-title":"Infect. Immun."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"e00242-21","DOI":"10.1128\/JB.00242-21","article-title":"Shigella flexneri Diguanylate Cyclases Regulate Virulence","volume":"203","author":"Ojha","year":"2021","journal-title":"J. Bacteriol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"387","DOI":"10.1146\/annurev-micro-020518-115555","article-title":"Multiple roles of c-di-GMP signaling in bacterial pathogenesis","volume":"73","author":"Valentini","year":"2019","journal-title":"Annu. Rev. Microbiol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1128\/MMBR.00043-12","article-title":"Cyclic di-GMP: The first 25 years of a universal bacterial second messenger","volume":"77","author":"Galperin","year":"2013","journal-title":"Microbiol. Mol. Biol. Rev."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"263","DOI":"10.1038\/nrmicro2109","article-title":"Principles of c-di-GMP signalling in bacteria","volume":"7","author":"Hengge","year":"2009","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"e1454","DOI":"10.1002\/wrna.1454","article-title":"Cyclic-di-GMP regulation of virulence in bacterial pathogens","volume":"9","author":"Hall","year":"2018","journal-title":"Wiley Interdiscip. Rev. RNA"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"385","DOI":"10.1146\/annurev.genet.40.110405.090423","article-title":"Mechanisms of cyclic-di-GMP signaling in bacteria","volume":"40","author":"Jenal","year":"2006","journal-title":"Annu. Rev. Genet."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"12547","DOI":"10.1074\/jbc.R115.711507","article-title":"Biofilms and cyclic di-GMP (c-di-GMP) signaling: Lessons from Pseudomonas aeruginosa and other bacteria","volume":"291","author":"Valentini","year":"2016","journal-title":"J. Biol. Chem."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"3.2.27","DOI":"10.1128\/microbiolspec.MB-0003-2014","article-title":"c-di-GMP and its effects on biofilm formation and dispersion: A Pseudomonas aeruginosa review","volume":"3","author":"Ha","year":"2015","journal-title":"Microbiol. Spectr."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Giardina, G., Paiardini, A., Fernicola, S., Franceschini, S., Rinaldo, S., Stelitano, V., and Cutruzzola, F. (2013). Investigating the allosteric regulation of YfiN from Pseudomonas aeruginosa: Clues from the structure of the catalytic domain. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0081324"},{"key":"ref_29","first-page":"1","article-title":"Structural insights into YfiR sequestering by YfiB in Pseudomonas aeruginosa PAO1","volume":"5","author":"Li","year":"2015","journal-title":"Sci. Rep."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"997","DOI":"10.1016\/j.bbrc.2018.10.190","article-title":"Structural analysis of activating mutants of YfiB from Pseudomonas aeruginosa PAO1","volume":"506","author":"Li","year":"2018","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Malone, J.G., Jaeger, T., Spangler, C., Ritz, D., Spang, A., Arrieumerlou, C., and Jenal, U. (2010). YfiBNR mediates cyclic di-GMP dependent small colony variant formation and persistence in Pseudomonas aeruginosa. PLoS Pathog., 6.","DOI":"10.1371\/journal.ppat.1000804"},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Malone, J.G., Jaeger, T., Manfredi, P., D\u00f6tsch, A., Blanka, A., Bos, R., Cornelis, G.R., H\u00e4ussler, S., and Jenal, U. (2012). The YfiBNR signal transduction mechanism reveals novel targets for the evolution of persistent Pseudomonas aeruginosa in cystic fibrosis airways. PLoS Pathog., 8.","DOI":"10.1371\/journal.ppat.1002760"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"471","DOI":"10.1111\/j.1432-1033.1988.tb14304.x","article-title":"Structural and immunochemical studies of the lipopolysaccharide from a new provisional serotype of Shigella flexneri","volume":"176","author":"WEHLER","year":"1988","journal-title":"Eur. J. Biochem."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"6640","DOI":"10.1073\/pnas.120163297","article-title":"One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products","volume":"97","author":"Datsenko","year":"2000","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/1471-2199-4-11","article-title":"Lambda Red-mediated recombinogenic engineering of enterohemorrhagic and enteropathogenic E. coli","volume":"4","author":"Murphy","year":"2003","journal-title":"BMC Mol. Biol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"996","DOI":"10.1128\/jcm.22.6.996-1006.1985","article-title":"Adherence of coagulase-negative staphylococci to plastic tissue culture plates: A quantitative model for the adherence of staphylococci to medical devices","volume":"22","author":"Christensen","year":"1985","journal-title":"J. Clin. Microbiol."},{"key":"ref_37","first-page":"e57322","article-title":"Bile salt-induced biofilm formation in enteric pathogens: Techniques for identification and quantification","volume":"135","author":"Nickerson","year":"2018","journal-title":"J. Vis. Exp."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"e00387-21","DOI":"10.1128\/IAI.00387-21","article-title":"Biofilm Formation and Virulence of Shigella flexneri Are Modulated by pH of Gastrointestinal Tract","volume":"89","author":"Chiang","year":"2021","journal-title":"Infect. Immun."},{"key":"ref_39","first-page":"1821","article-title":"In vitro assay of bacterial adhesion onto mammalian epithelial cells","volume":"117","author":"Letourneau","year":"2018","journal-title":"J. Vis. Exp."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"e57","DOI":"10.1002\/cpmc.57","article-title":"Shigella pathogenesis modeling with tissue culture assays","volume":"50","author":"Koestler","year":"2018","journal-title":"Curr. Protoc. Microbiol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"e2178","DOI":"10.7717\/peerj.2178","article-title":"Characterization of a biofilm-forming Shigella flexneri phenotype due to deficiency in Hep biosynthesis","volume":"4","author":"Xu","year":"2016","journal-title":"PeerJ"},{"key":"ref_42","unstructured":"Omidi, S., Dallal, M.M.S., Davoodabadi, A., Fard, R.M.N., Usefi, M., and Bakhtiari, R. (2017). Invasion of HEp-2 cells by Shigella spp. isolated from acute pediatric diarrhea. GMS Infect. Dis., 5."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"e01777-18","DOI":"10.1128\/mBio.01777-18","article-title":"Formate promotes Shigella intercellular spread and virulence gene expression","volume":"9","author":"Koestler","year":"2018","journal-title":"mBio"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"124","DOI":"10.1128\/iai.48.1.124-129.1985","article-title":"Plaque formation by virulent Shigella flexneri","volume":"48","author":"Oaks","year":"1985","journal-title":"Infect. Immun."},{"key":"ref_45","first-page":"e3293","article-title":"Plaque Assay to Determine Invasion and Intercellular Dissemination of Shigella flexneri in TC7 Human Intestinal Epithelial Cells","volume":"9","author":"Sharma","year":"2019","journal-title":"Bio-Protocol"},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"George, D.T., Mathesius, U., Behm, C.A., and Verma, N.K. (2014). The periplasmic enzyme, AnsB, of Shigella flexneri modulates bacterial adherence to host epithelial cells. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0094954"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"100","DOI":"10.1016\/j.devcel.2019.03.010","article-title":"Lifespan extension in C. elegans caused by bacterial colonization of the intestine and subsequent activation of an innate immune response","volume":"49","author":"Kumar","year":"2019","journal-title":"Dev. Cell"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"5043","DOI":"10.1128\/AEM.00558-06","article-title":"The Caenorhabditis elegans ABL-1 tyrosine kinase is required for Shigella flexneri pathogenesis","volume":"72","author":"Burton","year":"2006","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_49","doi-asserted-by":"crossref","unstructured":"Sen, T., and Verma, N.K. (2020). Functional annotation and curation of hypothetical proteins present in a newly emerged serotype 1c of Shigella flexneri: Emphasis on selecting targets for virulence and vaccine design studies. Genes, 11.","DOI":"10.3390\/genes11030340"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"403","DOI":"10.1016\/S0022-2836(05)80360-2","article-title":"Basic local alignment search tool","volume":"215","author":"Altschul","year":"1990","journal-title":"J. Mol. Biol."},{"key":"ref_51","first-page":"D237","article-title":"CDD: A conserved domain database for interactive domain family analysis","volume":"35","author":"Anderson","year":"2006","journal-title":"Nucleic Acids Res."},{"key":"ref_52","doi-asserted-by":"crossref","unstructured":"Gasteiger, E., Hoogland, C., Gattiker, A., Duvaud, S., Wilkins, M.R., Appel, R.D., and Bairoch, A. (2005). Protein identification and analysis tools on the ExPASy server. The Proteomics Protocols Handbook, Springer.","DOI":"10.1385\/1-59259-890-0:571"},{"key":"ref_53","doi-asserted-by":"crossref","unstructured":"Yu, C.-S., Cheng, C.-W., Su, W.-C., Chang, S.-C., Huang, S.-W., Hwang, J.-K., and Lu, C.-H. (2014). CELLO2GO: A Web Server for Protein subCELlular LOcalization Prediction with Functional Gene Ontology Annotation. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0099368"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"1608","DOI":"10.1093\/bioinformatics\/btq249","article-title":"PSORTb 3.0: Improved protein subcellular localization prediction with refined localization subcategories and predictive capabilities for all prokaryotes","volume":"26","author":"Yu","year":"2010","journal-title":"Bioinformatics"},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"2522","DOI":"10.1093\/bioinformatics\/bti309","article-title":"PSLpred: Prediction of subcellular localization of bacterial proteins","volume":"21","author":"Bhasin","year":"2005","journal-title":"Bioinformatics"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"567","DOI":"10.1006\/jmbi.2000.4315","article-title":"Predicting transmembrane protein topology with a hidden markov model: Application to complete genomes11Edited by F. Cohen","volume":"305","author":"Krogh","year":"2001","journal-title":"J. Mol. Boil."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"849","DOI":"10.1093\/bioinformatics\/17.9.849","article-title":"The HMMTOP transmembrane topology prediction server","volume":"17","author":"Simon","year":"2001","journal-title":"Bioinformatics"},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"42","DOI":"10.1016\/S1672-0229(06)60015-6","article-title":"VICMpred: An SVM-based method for the prediction of functional proteins of Gram-negative bacteria using amino acid patterns and composition","volume":"4","author":"Saha","year":"2006","journal-title":"Genom. Proteom. Bioinform."},{"key":"ref_59","doi-asserted-by":"crossref","unstructured":"Garg, A., and Gupta, D. (2008). VirulentPred: A SVM based prediction method for virulent proteins in bacterial pathogens. BMC Bioinform., 9.","DOI":"10.1186\/1471-2105-9-62"},{"key":"ref_60","first-page":"2","article-title":"Multiple sequence alignment using ClustalW and ClustalX","volume":"1","author":"Thompson","year":"2003","journal-title":"Curr. Protoc. Bioinform."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"725","DOI":"10.1038\/nprot.2010.5","article-title":"I-TASSER: A unified platform for automated protein structure and function prediction","volume":"5","author":"Roy","year":"2010","journal-title":"Nat. Protoc."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"W174","DOI":"10.1093\/nar\/gkv342","article-title":"I-TASSER server: New development for protein structure and function predictions","volume":"43","author":"Yang","year":"2015","journal-title":"Nucleic Acids Res."},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"1188","DOI":"10.1101\/gr.849004","article-title":"WebLogo: A sequence logo generator","volume":"14","author":"Crooks","year":"2004","journal-title":"Genome Res."},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"3022","DOI":"10.1093\/molbev\/msab120","article-title":"MEGA11: Molecular Evolutionary Genetics Analysis version 11","volume":"38","author":"Tamura","year":"2021","journal-title":"Mol. Biol. Evol."},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"11030","DOI":"10.1073\/pnas.0404206101","article-title":"Kumar S Prospects for inferring very large phylogenies by using the neighbor-joining method","volume":"101","author":"Tamura","year":"2004","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_66","first-page":"5","article-title":"An overview of composite likelihood methods","volume":"21","author":"Varin","year":"2011","journal-title":"Stat. Sin."},{"key":"ref_67","doi-asserted-by":"crossref","unstructured":"Isono, E., and Schwechheimer, C. (2010). Co-immunoprecipitation and Protein Blots. Plant Developmental Biology, Humana Press.","DOI":"10.1007\/978-1-60761-765-5_25"},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"1503","DOI":"10.1128\/jb.88.5.1503-1518.1964","article-title":"Epithelial cell penetration as an essential step in the pathogenesis of bacillary dysentery","volume":"88","author":"Labrec","year":"1964","journal-title":"J. Bacteriol."},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"99","DOI":"10.3934\/molsci.2020007","article-title":"Shigella pathogenesis: Molecular and computational insights","volume":"7","author":"Mukhopadhyay","year":"2020","journal-title":"AIMS Mol. Sci."},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"651","DOI":"10.1007\/s00253-010-3074-5","article-title":"GGDEF proteins YeaI, YedQ, and YfiN reduce early biofilm formation and swimming motility in Escherichia coli","volume":"90","author":"Hu","year":"2011","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"3089","DOI":"10.1128\/IAI.01396-12","article-title":"Genetic analysis of the role of yfiR in the ability of Escherichia coli CFT073 to control cellular cyclic dimeric GMP levels and to persist in the urinary tract","volume":"81","author":"Raterman","year":"2013","journal-title":"Infect. Immun."},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"4576","DOI":"10.1016\/j.jmb.2020.06.006","article-title":"Local c-di-GMP signaling in the control of synthesis of the E. coli biofilm exopolysaccharide pEtN-cellulose","volume":"432","author":"Richter","year":"2020","journal-title":"J. Mol. Biol."},{"key":"ref_73","doi-asserted-by":"crossref","unstructured":"Zhang, Y., Xia, B., Li, M., Shi, J., Long, Y., Jin, Y., Bai, F., Cheng, Z., Jin, S., and Wu, W. (2018). HigB reciprocally controls biofilm formation and the expression of type III secretion system genes through influencing the intracellular c-di-GMP level in Pseudomonas aeruginosa. Toxins, 10.","DOI":"10.3390\/toxins10110424"},{"key":"ref_74","doi-asserted-by":"crossref","first-page":"8487","DOI":"10.1073\/pnas.0602138103","article-title":"Genetic adaptation by Pseudomonas aeruginosa to the airways of cystic fibrosis patients","volume":"103","author":"Smith","year":"2006","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_75","doi-asserted-by":"crossref","first-page":"e3370","DOI":"10.1093\/femsle\/fnu045","article-title":"Virulence variations in Shigella and enteroinvasive Escherichia coli using the Caenorhabditis elegans model","volume":"362","author":"Fung","year":"2015","journal-title":"FEMS Microbiol. Lett."},{"key":"ref_76","doi-asserted-by":"crossref","first-page":"1222","DOI":"10.1128\/IAI.01077-13","article-title":"Three antagonistic cyclic di-GMP-catabolizing enzymes promote differential Dot\/Icm effector delivery and intracellular survival at the early steps of Legionella pneumophila infection","volume":"82","author":"Allombert","year":"2014","journal-title":"Infect. Immun."},{"key":"ref_77","doi-asserted-by":"crossref","first-page":"6335","DOI":"10.1073\/pnas.1901051116","article-title":"Cyclic-di-GMP regulation promotes survival of a slow-replicating subpopulation of intracellular Salmonella Typhimurium","volume":"116","author":"Petersen","year":"2019","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_78","first-page":"1","article-title":"Terrein is an inhibitor of quorum sensing and c-di-GMP in Pseudomonas aeruginosa: A connection between quorum sensing and c-di-GMP","volume":"8","author":"Kim","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_79","doi-asserted-by":"crossref","first-page":"1339","DOI":"10.3389\/fmicb.2021.675843","article-title":"Strategies for interfering with bacterial early stage biofilms","volume":"12","author":"Fu","year":"2021","journal-title":"Front. Microbiol."},{"key":"ref_80","doi-asserted-by":"crossref","first-page":"419","DOI":"10.1007\/978-1-4939-7240-1_31","article-title":"Targeting c-di-GMP signaling, biofilm formation, and bacterial motility with small molecules","volume":"1657","author":"Sintim","year":"2017","journal-title":"C-Di-GMP Signal."}],"container-title":["Microorganisms"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2607\/10\/3\/653\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T22:39:05Z","timestamp":1760135945000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2607\/10\/3\/653"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,3,18]]},"references-count":80,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2022,3]]}},"alternative-id":["microorganisms10030653"],"URL":"https:\/\/doi.org\/10.3390\/microorganisms10030653","relation":{"has-preprint":[{"id-type":"doi","id":"10.1101\/2021.09.20.461158","asserted-by":"object"}]},"ISSN":["2076-2607"],"issn-type":[{"value":"2076-2607","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,3,18]]}}}