Active Transport of Phosphorylated Carbohydrates Promotes Intestinal Colonization and Transmission of a Bacterial Pathogen
Essentially all Gram-negative pathogens are reliant on specific transport machineries termed binding protein-dependent transporters (BPDTs) to transport solutes such as amino acids, sugars and metal ions across their membranes. In this study we investigated AfuABC, a predicted iron-transporting BPDT found in many bacterial pathogens. We show by structural and functional approaches that AfuABC is not an iron transporter. Instead, AfuABC is a trio of proteins that bind and transport sugar-phosphates such as glucose-6-phosphate (G6P). In doing so, we present the first structural solution of a G6P-specific transport protein and add to the few known unique machineries for sugar-phosphate uptake by bacteria. Furthermore, we show that AfuABC is required by the intestinal pathogen C. rodentium to effectively transmit between mice and re-establish infection, leading us to propose that the transport of sugar-phosphates is an important part of general bacterial pathogenesis.
Vyšlo v časopise:
Active Transport of Phosphorylated Carbohydrates Promotes Intestinal Colonization and Transmission of a Bacterial Pathogen. PLoS Pathog 11(8): e32767. doi:10.1371/journal.ppat.1005107
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.ppat.1005107
Souhrn
Essentially all Gram-negative pathogens are reliant on specific transport machineries termed binding protein-dependent transporters (BPDTs) to transport solutes such as amino acids, sugars and metal ions across their membranes. In this study we investigated AfuABC, a predicted iron-transporting BPDT found in many bacterial pathogens. We show by structural and functional approaches that AfuABC is not an iron transporter. Instead, AfuABC is a trio of proteins that bind and transport sugar-phosphates such as glucose-6-phosphate (G6P). In doing so, we present the first structural solution of a G6P-specific transport protein and add to the few known unique machineries for sugar-phosphate uptake by bacteria. Furthermore, we show that AfuABC is required by the intestinal pathogen C. rodentium to effectively transmit between mice and re-establish infection, leading us to propose that the transport of sugar-phosphates is an important part of general bacterial pathogenesis.
Zdroje
1. Rohmer L, Hocquet D, Miller SI (2011) Are pathogenic bacteria just looking for food? Metabolism and microbial pathogenesis. Trends Microbiol 19: 341–348. doi: 10.1016/j.tim.2011.04.003 21600774
2. Hollenstein K, Dawson RJP, Locher KP (2007) Structure and mechanism of ABC transporter proteins. Curr Opin Struct Biol 17: 412–418. doi: 10.1016/j.sbi.2007.07.003 17723295
3. Chin N, Frey J, Chang CF, Chang YF (1996) Identification of a locus involved in the utilization of iron by Actinobacillus pleuropneumoniae. FEMS Microbiol Lett 143: 1–6. 8807793
4. Moisi M, Lichtenegger S, Tutz S, Seper A, Schild S, et al. (2013) Characterizing the hexose-6-phosphate transport system of Vibrio cholerae, a utilization system for carbon and phosphate sources. J Bacteriol 195: 1800–1808. doi: 10.1128/JB.01952-12 23417487
5. Hsu Y-M, Chin N, Chang C-F, Chang Y-F (2003) Cloning and characterization of the Actinobacillus pleuropneumoniae fur gene and its role in regulation of ApxI and AfuABC expression. DNA Seq 14: 169–181. doi: 10.1080/1042517031000089469 14509829
6. Wyckoff EE, Mey AR, Leimbach A, Fisher CF, Payne SM (2006) Characterization of ferric and ferrous iron transport systems in Vibrio cholerae. J Bacteriol 188: 6515–6523. doi: 10.1128/JB.00626-06 16952942
7. Klitgaard K, Friis C, Jensen TK, Angen Ø, Boye M (2012) Transcriptional portrait of Actinobacillus pleuropneumoniae during acute disease—potential strategies for survival and persistence in the host. PLoS One 7: e35549. doi: 10.1371/journal.pone.0035549 22530048
8. Whitby PW, Vanwagoner TM, Seale TW, Morton DJ, Stull TL (2006) Transcriptional profile of Haemophilus influenzae: effects of iron and heme. J Bacteriol 188: 5640–5645. doi: 10.1128/JB.00417-06 16855256
9. Bergholz TM, Wick LM, Qi W, Riordan JT, Ouellette LM, et al. (2007) Global transcriptional response of Escherichia coli O157:H7 to growth transitions in glucose minimal medium. BMC Microbiol 7: 97. doi: 10.1186/1471-2180-7-97 17967175
10. Berntsson RP-A, Smits SHJ, Schmitt L, Slotboom D-J, Poolman B (2010) A structural classification of substrate-binding proteins. FEBS Lett 584: 2606–2617. doi: 10.1016/j.febslet.2010.04.043 20412802
11. Clasquin MF, Melamud E, Singer A, Gooding JR, Xu X, et al. (2011) Riboneogenesis in yeast. Cell 145: 969–980. doi: 10.1016/j.cell.2011.05.022 21663798
12. Hall JA, Maloney PC (2005) Altered oxyanion selectivity in mutants of UhpT, the Pi-linked sugar phosphate carrier of Escherichia coli. J Biol Chem 280: 3376–3381. doi: 10.1074/jbc.M409965200 15556940
13. Weston LA, Kadner RJ (1987) Identification of uhp polypeptides and evidence for their role in exogenous induction of the sugar phosphate transport system of Escherichia coli K-12. J Bacteriol 169: 3546–3555. 3038843
14. Götz A, Goebel W (2010) Glucose and glucose 6-phosphate as carbon sources in extra- and intracellular growth of enteroinvasive Escherichia coli and Salmonella enterica. Microbiology 156: 1176–1187. doi: 10.1099/mic.0.034744–0 20075042
15. Collins JW, Keeney KM, Crepin VF, Rathinam VAK, Fitzgerald KA, et al. (2014) Citrobacter rodentium: infection, inflammation and the microbiota. Nat Rev Microbiol 12: 612–623. doi: 10.1038/nrmicro3315 25088150
16. Pacheco AR, Curtis MM, Ritchie JM, Munera D, Waldor MK, et al. (2012) Fucose sensing regulates bacterial intestinal colonization. Nature 492: 113–117. doi: 10.1038/nature11623 23160491
17. Perna NT, Plunkett G, Burland V, Mau B, Glasner JD, et al. (2001) Genome sequence of enterohaemorrhagic Escherichia coli O157:H7. Nature 409: 529–533. doi: 10.1038/35054089 11206551
18. Petty NK, Bulgin R, Crepin VF, Cerdeño-Tárraga AM, Schroeder GN, et al. (2010) The Citrobacter rodentium genome sequence reveals convergent evolution with human pathogenic Escherichia coli. J Bacteriol 192: 525–538. doi: 10.1128/JB.01144-09 19897651
19. Ma C, Wickham ME, Guttman JA, Deng W, Walker J, et al. (2006) Citrobacter rodentium infection causes both mitochondrial dysfunction and intestinal epithelial barrier disruption in vivo: role of mitochondrial associated protein (Map). Cell Microbiol 8: 1669–1686. doi: 10.1111/j.1462-5822.2006.00741.x 16759225
20. Wickham ME, Lupp C, Vázquez A, Mascarenhas M, Coburn B, et al. (2007) Citrobacter rodentium virulence in mice associates with bacterial load and the type III effector NleE. Microbes Infect 9: 400–407. doi: 10.1016/j.micinf.2006.12.016 17317262
21. Kelly M, Hart E, Mundy R, Marchès O, Wiles S, et al. (2006) Essential role of the type III secretion system effector NleB in colonization of mice by Citrobacter rodentium. Infect Immun 74: 2328–2337. doi: 10.1128/IAI.74.4.2328–2337.2006 16552063
22. Theriot CM, Koenigsknecht MJ, Carlson PE, Hatton GE, Nelson AM, et al. (2014) Antibiotic-induced shifts in the mouse gut microbiome and metabolome increase susceptibility to Clostridium difficile infection. Nat Commun 5: 3114. doi: 10.1038/ncomms4114 24445449
23. Antunes LCM, McDonald JAK, Schroeter K, Carlucci C, Ferreira RBR, et al. (2014) Antivirulence activity of the human gut metabolome. MBio 5: e01183–14. doi: 10.1128/mBio.01183-14 25073640
24. Ratledge C, Dover L (2000) Iron metabolism in pathogenic bacteria. Annu Rev Microbiol: 881–941. 11018148
25. Chico-Calero I, Suárez M, González-Zorn B, Scortti M, Slaghuis J, et al. (2002) Hpt, a bacterial homolog of the microsomal glucose- 6-phosphate translocase, mediates rapid intracellular proliferation in Listeria. Proc Natl Acad Sci U S A 99: 431–436. doi: 10.1073/pnas.012363899 11756655
26. Schwoppe C, Winkler HH, Neuhaus HE (2002) Properties of the Glucose-6-Phosphate Transporter from Chlamydia pneumoniae (HPTcp) and the Glucose-6-Phosphate Sensor from Escherichia coli (UhpC). J Bacteriol 184: 2108–2115. doi: 10.1128/JB.184.8.2108–2115.2002 11914341
27. Engström P, Krishnan KS, Ngyuen BD, Chorell E, Normark J, et al. (2015) A 2-Pyridone-Amide Inhibitor Targets the Glucose Metabolism Pathway of Chlamydia trachomatis. MBio 6: 1–9. doi: 10.1128/mBio.02304-14.Editor
28. Gaudet RG, Sintsova A, Buckwalter CM, Leung N, Cochrane A, et al. (2015) Cytosolic detection of the bacterial metabolite HBP activates TIFA-dependent innate immunity. Science 348: 1251–1255. doi: 10.1126/science.aaa4921 26068852
29. Sham HP, Yu EYS, Gulen MF, Bhinder G, Stahl M, et al. (2013) SIGIRR, a Negative Regulator of TLR/IL-1R Signalling Promotes Microbiota Dependent Resistance to Colonization by Enteric Bacterial Pathogens. PLoS Pathog 9. doi: 10.1371/journal.ppat.1003539
30. Ng KM, Ferreyra JA, Higginbottom SK, Lynch JB, Kashyap PC, et al. (2013) Microbiota-liberated host sugars facilitate post-antibiotic expansion of enteric pathogens. Nature 502: 96–99. doi: 10.1038/nature12503 23995682
31. Siegel SJ, Roche AM, Weiser JN (2014) Influenza promotes pneumococcal growth during coinfection by providing host sialylated substrates as a nutrient source. Cell Host Microbe 16: 55–67. doi: 10.1016/j.chom.2014.06.005 25011108
32. Holmén Larsson JM, Thomsson KA, Rodríguez-Piñeiro AM, Karlsson H, Hansson GC (2013) Studies of mucus in mouse stomach, small intestine, and colon. III. Gastrointestinal Muc5ac and Muc2 mucin O-glycan patterns reveal a regiospecific distribution. Am J Physiol Gastrointest Liver Physiol 305: G357–G363. doi: 10.1152/ajpgi.00048.2013 23832516
33. Bai J, McAteer SP, Paxton E, Mahajan A, Gally DL, et al. (2011) Screening of an E. coli O157:H7 Bacterial Artificial Chromosome Library by Comparative Genomic Hybridization to Identify Genomic Regions Contributing to Growth in Bovine Gastrointestinal Mucus and Epithelial Cell Colonization. Front Microbiol 2: 168. doi: 10.3389/fmicb.2011.00168 21887152
34. Bergstrom KSB, Kissoon-Singh V, Gibson DL, Ma C, Montero M, et al. (2010) Muc2 protects against lethal infectious colitis by disassociating pathogenic and commensal bacteria from the colonic mucosa. PLoS Pathog 6. doi: 10.1371/journal.ppat.1000902
35. Ulrich A, Andersen KR, Schwartz TU (2012) Exponential megapriming PCR (EMP) cloning—seamless DNA insertion into any target plasmid without sequence constraints. PLoS One 7: e53360. doi: 10.1371/journal.pone.0053360 23300917
36. van den Ent F, Löwe J (2006) RF cloning: a restriction-free method for inserting target genes into plasmids. J Biochem Biophys Methods 67: 67–74. doi: 10.1016/j.jbbm.2005.12.008 16480772
37. Adams PD, Afonine P V, Bunkóczi G, Chen VB, Echols N, et al. (2011) The Phenix software for automated determination of macromolecular structures. Methods 55: 94–106. doi: 10.1016/j.ymeth.2011.07.005 21821126
38. Emsley P, Lohkamp B, Scott WG, Cowtan K (2010) Features and development of Coot. Acta Crystallogr D Biol Crystallogr 66: 486–501. doi: 10.1107/S0907444910007493 20383002
39. Pettersen EF, Goddard TD, Huang CC, Couch GS, Greenblatt DM, et al. (2004) UCSF Chimera—a visualization system for exploratory research and analysis. J Comput Chem 25: 1605–1612. doi: 10.1002/jcc.20084 15264254
40. Baba T, Ara T, Hasegawa M, Takai Y, Okumura Y, et al. (2006) Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection. Mol Syst Biol 2: 2006.0008. doi: 10.1038/msb4100050
41. Edgar RC (2004) MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Res 32: 1792–1797. doi: 10.1093/nar/gkh340 15034147
42. Ronquist F, Huelsenbeck JP (2003) MrBayes 3: Bayesian phylogenetic inference under mixed models. Bioinformatics 19: 1572–1574. doi: 10.1093/bioinformatics/btg180 12912839
43. Edwards RA, Keller LH, Schifferli DM (1998) Improved allelic exchange vectors and their use to analyze 987P fimbria gene expression. Gene 207: 149–157. doi: 10.1016/S0378-1119(97)00619-7 9511756
44. Wickham ME, Brown NF, Boyle EC, Coombes BK, Finlay BB (2007) Virulence Is Positively Selected by Transmission Success between Mammalian Hosts. Curr Biol 17: 783–788. doi: 10.1016/j.cub.2007.03.067 17442572
45. Kenny B, Abe A, Stein M, Finlay BB (1997) Enteropathogenic Escherichia coli protein secretion is induced in response to conditions similar to those in the gastrointestinal tract. Infect Immun 65: 2606–2612. 9199427
46. Newman J V, Zabel BA, Jha SS, Schauer DB (1999) Citrobacter rodentium espB is necessary for signal transduction and for infection of laboratory mice. Infect Immun 67: 6019–6025. 10531262
47. Knodler LA, Crowley SM, Sham HP, Yang H, Wrande M, et al. (2014) Noncanonical Inflammasome Activation of Caspase-4/Caspase-11 Mediates Epithelial Defenses against Enteric Bacterial Pathogens. Cell Host Microbe 16: 249–256. doi: 10.1016/j.chom.2014.07.002 25121752
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Hygiena a epidemiológia Infekčné lekárstvo LaboratóriumČlánok vyšiel v časopise
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