Evolution of Multidrug Resistance during Infection Involves Mutation of the Essential Two Component Regulator WalKR
Antimicrobial resistance in Staphylococcus aureus is a major public health threat, compounded by emergence of strains with resistance to vancomycin and daptomycin, both last line antimicrobials. Here we have performed high throughput DNA sequencing and comparative genomics for five clinical pairs of vancomycin-susceptible (VSSA) and vancomycin-intermediate ST239 S. aureus (VISA); each pair isolated before and after vancomycin treatment failure. These comparisons revealed a frequent pattern of mutation among the VISA strains within the essential walKR two-component regulatory locus involved in control of cell wall metabolism. We then conducted bi-directional allelic exchange experiments in our clinical VSSA and VISA strains and showed that single nucleotide substitutions within either walK or walR lead to co-resistance to vancomycin and daptomycin, and caused the typical cell wall thickening observed in resistant clinical isolates. Ion Torrent genome sequencing confirmed no additional regulatory mutations had been introduced into either the walR or walK VISA mutants during the allelic exchange process. However, two potential compensatory mutations were detected within putative transport genes for the walK mutant. The minimal genetic changes in either walK or walR also attenuated virulence, reduced biofilm formation, and led to consistent transcriptional changes that suggest an important role for this regulator in control of central metabolism. This study highlights the dramatic impacts of single mutations that arise during persistent S. aureus infections and demonstrates the role played by walKR to increase drug resistance, control metabolism and alter the virulence potential of this pathogen.
Vyšlo v časopise:
Evolution of Multidrug Resistance during Infection Involves Mutation of the Essential Two Component Regulator WalKR. PLoS Pathog 7(11): e32767. doi:10.1371/journal.ppat.1002359
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.ppat.1002359
Souhrn
Antimicrobial resistance in Staphylococcus aureus is a major public health threat, compounded by emergence of strains with resistance to vancomycin and daptomycin, both last line antimicrobials. Here we have performed high throughput DNA sequencing and comparative genomics for five clinical pairs of vancomycin-susceptible (VSSA) and vancomycin-intermediate ST239 S. aureus (VISA); each pair isolated before and after vancomycin treatment failure. These comparisons revealed a frequent pattern of mutation among the VISA strains within the essential walKR two-component regulatory locus involved in control of cell wall metabolism. We then conducted bi-directional allelic exchange experiments in our clinical VSSA and VISA strains and showed that single nucleotide substitutions within either walK or walR lead to co-resistance to vancomycin and daptomycin, and caused the typical cell wall thickening observed in resistant clinical isolates. Ion Torrent genome sequencing confirmed no additional regulatory mutations had been introduced into either the walR or walK VISA mutants during the allelic exchange process. However, two potential compensatory mutations were detected within putative transport genes for the walK mutant. The minimal genetic changes in either walK or walR also attenuated virulence, reduced biofilm formation, and led to consistent transcriptional changes that suggest an important role for this regulator in control of central metabolism. This study highlights the dramatic impacts of single mutations that arise during persistent S. aureus infections and demonstrates the role played by walKR to increase drug resistance, control metabolism and alter the virulence potential of this pathogen.
Zdroje
1. WardPBJohnsonPDGrabschEAMayallBCGraysonML 2001 Treatment failure due to methicillin-resistant Staphylococcus aureus (MRSA) with reduced susceptibility to vancomycin. Med J Aust 175 480 483
2. HowdenBPWardPBCharlesPGKormanTMFullerA 2004 Treatment outcomes for serious infections caused by methicillin-resistant Staphylococcus aureus with reduced vancomycin susceptibility. Clin Infect Dis 38 521 528
3. HowdenBPDaviesJKJohnsonPDStinearTPGraysonML 2010 Reduced vancomycin susceptibility in Staphylococcus aureus, including vancomycin-intermediate and heterogeneous vancomycin-intermediate strains: resistance mechanisms, laboratory detection, and clinical implications. Clin Microbiol Rev 23 99 139
4. HowdenBPSmithDJMansellAJohnsonPDWardPB 2008 Different bacterial gene expression patterns and attenuated host immune responses are associated with the evolution of low-level vancomycin resistance during persistent methicillin-resistant Staphylococcus aureus bacteraemia. BMC Microbiol 8 39
5. HiramatsuKAritakaNHanakiHKawasakiSHosodaY 1997 Dissemination in Japanese hospitals of strains of Staphylococcus aureus heterogeneously resistant to vancomycin. Lancet 350 1670 1673
6. SieradzkiKRobertsRBHaberSWTomaszA 1999 The development of vancomycin resistance in a patient with methicillin-resistant Staphylococcus aureus infection. N Engl J Med 340 517 523
7. SieradzkiKTomaszA 2003 Alterations of cell wall structure and metabolism accompany reduced susceptibility to vancomycin in an isogenic series of clinical isolates of Staphylococcus aureus. J Bacteriol 185 7103 7110
8. SmithTLPearsonMLWilcoxKRCruzCLancasterMV 1999 Emergence of vancomycin resistance in Staphylococcus aureus. Glycopeptide-Intermediate Staphylococcus aureus Working Group. N Engl J Med 340 493 501
9. MooreMRPerdreau-RemingtonFChambersHF 2003 Vancomycin treatment failure associated with heterogeneous vancomycin-intermediate Staphylococcus aureus in a patient with endocarditis and in the rabbit model of endocarditis. Antimicrob Agents Chemother 47 1262 1266
10. KelleyPGGaoWWardPBHowdenBP 2011 Daptomycin non-susceptibility in vancomycin-intermediate Staphylococcus aureus (VISA) and heterogeneous-VISA (hVISA): implications for therapy after vancomycin treatment failure. J Antimicrob Chemother 66 1057 1060
11. MwangiMMWuSWZhouYSieradzkiKde LencastreH 2007 Tracking the in vivo evolution of multidrug resistance in Staphylococcus aureus by whole-genome sequencing. Proc Natl Acad Sci U S A 104 9451 9456
12. NeohHMCuiLYuzawaHTakeuchiFMatsuoM 2008 Mutated response regulator graR is responsible for phenotypic conversion of Staphylococcus aureus from heterogeneous vancomycin-intermediate resistance to vancomycin-intermediate resistance. Antimicrob Agents Chemother 52 45 53
13. CuiLNeohHMShojiMHiramatsuK 2009 Contribution of vraSR and graSR point mutations to vancomycin resistance in vancomycin-intermediate Staphylococcus aureus. Antimicrob Agents Chemother 53 1231 1234
14. CuiLMurakamiHKuwahara-AraiKHanakiHHiramatsuK 2000 Contribution of a thickened cell wall and its glutamine nonamidated component to the vancomycin resistance expressed by Staphylococcus aureus Mu50. Antimicrob Agents Chemother 44 2276 2285
15. HowdenBPJohnsonPDWardPBStinearTPDaviesJK 2006 Isolates with low-level vancomycin resistance associated with persistent methicillin-resistant Staphylococcus aureus bacteremia. Antimicrob Agents Chemother 50 3039 3047
16. RenzoniABarrasCFrancoisPCharbonnierYHugglerE 2006 Transcriptomic and functional analysis of an autolysis-deficient, teicoplanin-resistant derivative of methicillin-resistant Staphylococcus aureus. Antimicrob Agents Chemother 50 3048 3061
17. CuiLIsiiTFukudaMOchiaiTNeohHM 2010 An RpoB mutation confers dual heteroresistance to daptomycin and vancomycin in Staphylococcus aureus. Antimicrob Agents Chemother 54 5222 5233
18. HowdenBPStinearTPAllenDLJohnsonPDWardPB 2008 Genomic analysis reveals a point mutation in the two-component sensor gene graS that leads to intermediate vancomycin resistance in clinical Staphylococcus aureus. Antimicrob Agents Chemother 52 3755 3762
19. MoisePANorthDSteenbergenJNSakoulasG 2009 Susceptibility relationship between vancomycin and daptomycin in Staphylococcus aureus: facts and assumptions. Lancet Infect Dis 9 617 624
20. FriedmanLAlderJDSilvermanJA 2006 Genetic changes that correlate with reduced susceptibility to daptomycin in Staphylococcus aureus. Antimicrob Agents Chemother 50 2137 2145
21. HowdenBPSeemannTHarrisonPFMcEvoyCRStantonJA 2010 Complete genome sequence of Staphylococcus aureus strain JKD6008, an ST239 clone of methicillin-resistant Staphylococcus aureus with intermediate-level vancomycin resistance. J Bacteriol 192 5848 5849
22. OkadaAIgarashiMOkajimaTKinoshitaNUmekitaM 2010 Walkmycin B targets WalK (YycG), a histidine kinase essential for bacterial cell growth. J Antibiot (Tokyo) 63 89 94
23. QinZLeeBYangLZhangJYangX 2007 Antimicrobial activities of YycG histidine kinase inhibitors against Staphylococcus epidermidis biofilms. FEMS Microbiol Lett 273 149 156
24. GotohYDoiAFurutaEDubracSIshizakiY 2010 Novel antibacterial compounds specifically targeting the essential WalR response regulator. J Antibiot (Tokyo) 63 127 134
25. DubracSBisicchiaPDevineKMMsadekT 2008 A matter of life and death: cell wall homeostasis and the WalKR (YycGF) essential signal transduction pathway. Mol Microbiol 70 1307 1322
26. BourretRB 2010 Receiver domain structure and function in response regulator proteins. Curr Opin Microbiol 13 142 149
27. DoiAOkajimaTGotohYTanizawaKUtsumiR 2010 X-ray crystal structure of the DNA-binding domain of response regulator WalR essential to the cell viability of Staphylococcus aureus and interaction with target DNA. Biosci Biotechnol Biochem 74 1901 1907
28. HulkoMBerndtFGruberMLinderJUTruffaultV 2006 The HAMP domain structure implies helix rotation in transmembrane signaling. Cell 126 929 940
29. RepikARebbapragadaAJohnsonMSHaznedarJOZhulinIB 2000 PAS domain residues involved in signal transduction by the Aer redox sensor of Escherichia coli. Mol Microbiol 36 806 816
30. PelegAYMongaDPillaiSMylonakisEMoelleringRCJr 2009 Reduced susceptibility to vancomycin influences pathogenicity in Staphylococcus aureus infection. J Infect Dis 199 532 536
31. DubracSMsadekT 2004 Identification of genes controlled by the essential YycG/YycF two-component system of Staphylococcus aureus. J Bacteriol 186 1175 1181
32. DubracSBonecaIGPoupelOMsadekT 2007 New insights into the WalK/WalR (YycG/YycF) essential signal transduction pathway reveal a major role in controlling cell wall metabolism and biofilm formation in Staphylococcus aureus. J Bacteriol 189 8257 8269
33. KanehisaM 2002 The KEGG database. Novartis Found Symp 247: 91-101; discussion 101-103, 119-128, 244-152
34. KurodaMKurodaHOshimaTTakeuchiFMoriH 2003 Two-component system VraSR positively modulates the regulation of cell-wall biosynthesis pathway in Staphylococcus aureus. Mol Microbiol 49 807 821
35. McAleeseFWuSWSieradzkiKDunmanPMurphyE 2006 Overexpression of genes of the cell wall stimulon in clinical isolates of Staphylococcus aureus exhibiting vancomycin-intermediate- S. aureus-type resistance to vancomycin. J Bacteriol 188 1120 1133
36. McCallumNSpeharGBischoffMBerger-BachiB 2006 Strain dependence of the cell wall-damage induced stimulon in Staphylococcus aureus. Biochim Biophys Acta 1760 1475 1481
37. SobralRGJonesAEDes EtagesSGDoughertyTJPeitzschRM 2007 Extensive and genome-wide changes in the transcription profile of Staphylococcus aureus induced by modulating the transcription of the cell wall synthesis gene murF. J Bacteriol 189 2376 2391
38. UtaidaSDunmanPMMacapagalDMurphyEProjanSJ 2003 Genome-wide transcriptional profiling of the response of Staphylococcus aureus to cell-wall-active antibiotics reveals a cell-wall-stress stimulon. Microbiology 149 2719 2732
39. MartinPKLiTSunDBiekDPSchmidMB 1999 Role in cell permeability of an essential two-component system in Staphylococcus aureus. J Bacteriol 181 3666 3673
40. DelauneAPoupelOMalletACoicYMMsadekT 2011 Peptidoglycan crosslinking relaxation plays an important role in Staphylococcus aureus WalKR-dependent cell viability. PLoS One 6 e17054
41. ShojiMCuiLIizukaRKomotoANeohHM 2011 walK and clpP Mutations Confer Reduced Vancomycin Susceptibility in Staphylococcus aureus. Antimicrob Agents Chemother 55 3870 3881
42. WatanabeYCuiLKatayamaYKozueKHirakawaH 2011 Impact of rpoB mutations on reduced vancomycin susceptibility in Staphylococcus aureus. J Clin Microbiol 49 2680 2684
43. JansenATurckMSzekatCNagelMCleverI 2007 Role of insertion elements and yycFG in the development of decreased susceptibility to vancomycin in Staphylococcus aureus. Int J Med Microbiol 297 205 215
44. GaoWChuaKDaviesJKNewtonHJSeemannT 2010 Two novel point mutations in clinical Staphylococcus aureus reduce linezolid susceptibility and switch on the stringent response to promote persistent infection. PLoS Pathog 6 e1000944
45. SadykovMRMattesTALuongTTZhuYDaySR 2010 Tricarboxylic acid cycle-dependent synthesis of Staphylococcus aureus Type 5 and 8 capsular polysaccharides. J Bacteriol 192 1459 1462
46. BiswasRVogguLSimonUKHentschelPThummG 2006 Activity of the major staphylococcal autolysin Atl. FEMS Microbiol Lett 259 260 268
47. BolesBRThoendelMRothAJHorswillAR 2010 Identification of genes involved in polysaccharide-independent Staphylococcus aureus biofilm formation. PLoS One 5 e10146
48. MannEERiceKCBolesBREndresJLRanjitD 2009 Modulation of eDNA release and degradation affects Staphylococcus aureus biofilm maturation. PLoS One 4 e5822
49. NovickRP 2003 Autoinduction and signal transduction in the regulation of staphylococcal virulence. Mol Microbiol 48 1429 1449
50. WagnerCSaizieu AdASchonfeldHJKamberMLangeR 2002 Genetic analysis and functional characterization of the Streptococcus pneumoniae vic operon. Infect Immun 70 6121 6128
51. ThroupJPKoretkeKKBryantAPIngrahamKAChalkerAF 2000 A genomic analysis of two-component signal transduction in Streptococcus pneumoniae. Mol Microbiol 35 566 576
52. CLSI 2010 Performance Standards for Antimicrobial Susceptibility Testing. Approved Standard. CLSI document M100-S20 Wayne, , Pennsylvania CLSI; 2010
53. MoneckeSKanigHRudolphWMullerECoombsG 2010 Characterisation of Australian MRSA strains ST75- and ST883-MRSA-IV and analysis of their accessory gene regulator locus. PLoS One 5 e14025
54. O'BrienFGUdoEEGrubbWB 2006 Contour-clamped homogeneous electric field electrophoresis of Staphylococcus aureus. Nat Protoc 1 3028 3033
55. RothbergJMHinzWRearickTMSchultzJMileskiW 2011 An integrated semiconductor device enabling non-optical genome sequencing. Nature 475 348 352
56. RumbleSMLacroutePDalcaAVFiumeMSidowA 2009 SHRiMP: accurate mapping of short color-space reads. PLoS Comput Biol 5 e1000386
57. PieperUEswarNWebbBMEramianDKellyL 2009 MODBASE, a database of annotated comparative protein structure models and associated resources. Nucleic Acids Res 37 D347 354
58. SaalLHTroeinCVallon-ChristerssonJGruvbergerSBorgA 2002 BioArray Software Environment (BASE): a platform for comprehensive management and analysis of microarray data. Genome Biol 15 8
59. SmythGK 2004 Linear models and empirical bayes methods for assessing differential expression in microarray experiments. Stat Appl Genet Mol Biol 3 Article 3
60. BurianMRautenbergMKohlerTFritzMKrismerB 2010 Temporal expression of adhesion factors and activity of global regulators during establishment of Staphylococcus aureus nasal colonization. J Infect Dis 201 1414 1421
61. TatusovRLFedorovaNDJacksonJDJacobsARKiryutinB 2003 The COG database: an updated version includes eukaryotes. BMC Bioinformatics 4 41
62. PatronRLClimoMWGoldsteinBPArcherGL 1999 Lysostaphin treatment of experimental aortic valve endocarditis caused by a Staphylococcus aureus isolate with reduced susceptibility to vancomycin. Antimicrob Agents Chemother 43 1754 1755
63. KreiswirthBNLofdahlSBetleyMJO'ReillyMSchlievertPM 1983 The toxic shock syndrome exotoxin structural gene is not detectably transmitted by a prophage. Nature 305 709 712
64. DiepBAGillSRChangRFPhanTHChenJH 2006 Complete genome sequence of USA300, an epidemic clone of community-acquired meticillin-resistant Staphylococcus aureus. Lancet 367 731 739
65. BaeTSchneewindO 2006 Allelic replacement in Staphylococcus aureus with inducible counter-selection. Plasmid 55 58 63
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Hygiena a epidemiológia Infekčné lekárstvo LaboratóriumČlánok vyšiel v časopise
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