BMC Microbiology | |
Complete nucleotide sequence of pRS218, a large virulence plasmid, that augments pathogenic potential of meningitis-associated Escherichia coli strain RS218 | |
Subhashinie Kariyawasam2  Chitrita DebRoy1  Vivek Kapur1  Edward G Dudley2  Robab Katani1  Atul Chaudhari1  Kurundu Hewage Eranda M Karunathilake1  Dona Saumya S Wijetunge1  | |
[1] Department of Veterinary and Biomedical Sciences, Pennsylvania State University, University Park, 16802, PA, USA;Center for Molecular Immunology and Infectious Disease, University Park, 16802, PA, USA | |
关键词: Virulence; Plasmid; Meningitis; Neonates; Escherichia coli; DNA sequence; | |
Others : 1140478 DOI : 10.1186/s12866-014-0203-9 |
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received in 2014-03-17, accepted in 2014-07-18, 发布年份 2014 | |
【 摘 要 】
Background
Escherichia coli is the most predominant Gram-negative bacterial pathogen associated with neonatal meningitis. Previous studies indicated that the prototypic neonatal meningitis E. coli (NMEC) strain RS218 (O18:K1:H7) harbors one large plasmid. Objectives of the present study were to analyze the complete nucleotide sequence of this large plasmid (pRS218) and its contribution to NMEC pathogenesis using in vitro and in vivo models of neonatal meningitis.
Results
The plasmid is 114,231 bp in size, belongs to the incompatibility group FIB/IIA (IncFIB/IIA), and contains a genetic load region that encodes several virulence and fitness traits such as enterotoxicity, iron acquisition and copper tolerance. The nucleotide sequence of pRS218 showed a 41- 46% similarity to other neonatal meningitis-causing E. coli (NMEC) plasmids and remarkable nucleotide sequence similarity (up to 100%) to large virulence plasmids of E. coli associated with acute cystitis. Some genes located on pRS218 were overly represented by NMEC strains compared to fecal E. coli isolated from healthy individuals. The plasmid-cured strain was significantly attenuated relative to the RS218 wild-type strain as determined in vitro by invasion potential to human cerebral microvascular endothelial cells and in vivo by mortalities, histopathological lesions in the brain tissue, and bacterial recovery from the cerebrospinal fluid of infected rat pups.
Conclusions
The pRS218 is an IncFIB/IIA plasmid which shares a remarkable nucleotide sequence similarity to large plasmids of E. coli associated with cystitis. Both in vitro and in vivo experiments indicated that pRS218 plays an important role in NMEC pathogenesis.
【 授权许可】
2014 Wijetunge et al.; licensee BioMed Central Ltd.
【 预 览 】
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【 参考文献 】
- [1]Lawn JE, Cousens S, Zupan J: 4 million neonatal deaths: when? Where? Why? Lancet 2005, 365(9462):891-900.
- [2]Liu L, Johnson HL, Cousens S, Perin J, Scott S, Lawn JE, Rudan I, Campbell H, Cibulskis R, Li M, Mathers C, Black R: Global, regional, and national causes of child mortality: an updated systematic analysis for 2010 with time trends since 2000. Lancet 2012, 379(9832):2151-2161.
- [3]Bell AH, Brown D, Halliday HL, McClure G, McReid M: Meningitis in the newborn¿a 14 year review. Arch Dis Child 1989, 64(6):873-874.
- [4]Kim KS: Strategy of Escherichia coli for crossing the blood-brain barrier. J Infect Dis 2002, 186(Supplement 2):S220-S224.
- [5]Kim KS: Pathogenesis of bacterial meningitis: from bacteraemia to neuronal injury. Nat Rev Neurosci 2003, 4(5):376-385.
- [6]Frosch M, Edwards U, Bousset K, Krauße B, Weisgerber C: Evidence for a common molecular origin of the capsule gene loci in Gram-negative bacteria expressing group II capsular polysaccharides. Mol Microbiol 1991, 5(5):1251-1263.
- [7]Pong A, Bradley JS: Bacterial meningitis and the newborn infant. Infect Dis Clin North Am 1999, 13(3):711-733.
- [8]Polin RA, Harris MC: Neonatal bacterial meningitis. Sem Neonatol 2001, 6(2):157-172.
- [9]Jain R, Rivera MC, Moore JE, Lake JA: Horizontal gene transfer accelerates genome innovation and evolution. Mol Biolo Evol 2003, 20(10):1598-1602.
- [10]Johnson TJ, Nolan LK: Pathogenomics of the virulence plasmids of Escherichia coli. Microbiol Mol Biol Rev 2009, 73(4):750-774.
- [11]Carattoli A: Plasmids and the spread of resistance. Intl J Med Microbiol 2013, 303(6¿7):298-304.
- [12]Cusumano CK, Hung CS, Chen SL, Hultgren SJ: Virulence plasmid harbored by uropathogenic Escherichia coli functions in acute stages of pathogenesis. Infect Immun 2010, 78(4):1457-1467.
- [13]DebRoy C, Sidhu MS, Sarker U, Jayarao BM, Stell AL, Bell NP, Johnson TJ: Complete sequence of pEC14_114, a highly conserved IncFIB/FIIA plasmid associated with uropathogenic Escherichia coli cystitis strains. Plasmid 2010, 63(1):53-60.
- [14]Peigne C, Bidet P, Mahjoub-Messai F, Plainvert C, Barbe V, Médigue C, Frapy E, Nassif X, Denamur E, Bingen E, Bonacorsi S: The plasmid of Escherichia coli Strain S88 (O45:K1:H7) that causes neonatal meningitis is closely related to avian pathogenic E. coli plasmids and is associated with high-level bacteremia in a neonatal rat meningitis model. Infect Immun 2009, 77(6):2272-2284.
- [15]Lu S, Zhang X, Zhu Y, Kim KS, Yang J, Jin Q: Complete genome sequence of the neonatal-meningitis-associatedEscherichia colistrain CE10.J Bacteriol 2011, 193(24):7005.
- [16]Silver RP, Aaronson W, Vann WF: The K1 capsular polysaccharide of Escherichia coli. Rev Infect Dis 1988, 10(2):S282-S286.
- [17]Silver RP, Aaronson W, Sutton A, Schneerson R: Comparative analysis of plasmids and some metabolic characteristics of Escherichia coli K1 from diseased and healthy individuals. Infect Immun 1980, 29(1):200-206.
- [18]Tivendale KA, Logue CM, Kariyawasam S, Jordan D, Hussein A, Li G, Wannemuehler Y, Nolan LK: Avian-pathogenic Escherichia coli strains are similar to neonatal meningitis E. coli strains and are able to cause meningitis in the rat model of human disease. Infect Immun 2010, 78(8):3412-3419.
- [19]Verkhovsky MI, Bogachev AV, Pivtsov AV, Bertsova YV, Fedin MV, Bloch DA, Kulik LV: Sodium-dependent movement of covalently bound FMN Residue(s) in Na+-Translocating NADH: quinone oxidoreductase. Biochemistry 2012, 51(27):5414-5421.
- [20]Lehoux IE, Mazzulla MJ, Baker A, Petit CM: Purification and characterization of YihA, an essential GTP-binding protein from Escherichia coli. Protein Expr Purif 2003, 30(2):203-209.
- [21]Ripio MT, Brehm K, Lara M, Suárez M, Vázquez-Boland JA: Glucose-1-phosphate utilization by Listeria monocytogenes is PrfA dependent and coordinately expressed with virulence factors. J Bacteriol 1997, 179(22):7174-7180.
- [22]Scheurwater E, Reid CW, Clarke AJ: Lytic transglycosylases: bacterial space-making autolysins. Intl J Biochem Cell Biol 2008, 40(4):586-591.
- [23]Toh H, Oshima K, Toyoda A, Ogura Y, Ooka T, Sasamoto H, Park S-H, Iyoda S, Kurokawa K, Morita H, Itoh K, Taylor TD, Hayashi T, Hattori M: Complete genome sequence of the wild-type commensal Escherichia coli strain SE15, belonging to phylogenetic group B2. J Bacteriol 2010, 192(4):1165-1166.
- [24]Wajima T, Sabui S, Kano S, Ramamurthy T, Chatterjee NS, Hamabata T: Entire sequence of the colonization factor coli surface antigen 6-encoding plasmid pCss165 from an enterotoxigenic Escherichia coli clinical isolate. Plasmid 2013, 70(3):345-352.
- [25]Zhang W, Bielaszewska M, Kunsmann L, Mellmann A, Bauwens A, Köck R, Kossow A, Anders A, Gatermann S, Karch H: Lability of the pAA virulence plasmid inEscherichia coliO104:H4: implications for virulence in humans.PLoS One 2013, 8(6):e66717.
- [26]Logue CM, Doetkott C, Mangiamele P, Wannemuehler YM, Johnson TJ, Tivendale KA, Li G, Sherwood JS, Nolan LK: Genotypic and phenotypic traits that distinguish neonatal meningitis-associated Escherichia coli from fecal E. coli isolates of healthy human hosts. Appl Environ Microbiol 2012, 78(16):5824-5830.
- [27]Nakaya R, Nakamura A, Murata Y: Resistance transfer agents in Shigella. Biochem Biophys Res Commun 1960, 3:654-659.
- [28]Grass G, Rensing C, Solioz M: Metallic copper as an antimicrobial surface. Appl Environ Microbiol 2011, 77(5):1541-1547.
- [29]Gupta SD, Wu HC, Rick PD: A Salmonella typhimurium genetic locus which confers copper tolerance on copper-sensitive mutants of Escherichia coli. J Bacteriol 1997, 179(16):4977-4984.
- [30]Nataro JP, Seriwatana J, Fasano A, Maneval DR, Guers LD, Noriega F, Dubovsky F, Levine MM, Morris JG: Identification and cloning of a novel plasmid-encoded enterotoxin of enteroinvasive Escherichia coli and Shigella strains. Infect Immun 1995, 63(12):4721-4728.
- [31]Badger JL, Wass CA, Weissman SJ, Kim KS: Application of signature-tagged mutagenesis for identification of Escherichia coli K1 genes that contribute to invasion of human brain microvascular endothelial cells. Infect Immun 2000, 68(9):5056-5061.
- [32]Huang SH, Wass C, Fu Q, Prasadarao NV, Stins M, Kim KS: Escherichia coli invasion of brain microvascular endothelial cells in vitro and in vivo: molecular cloning and characterization of invasion gene ibe10. Infec Immun 1995, 63(11):4470-4475.
- [33]Sambrook J, Russell DW: Molecular cloning: a laboratory manual. In, Volume 1. 4th edition. New York: Cold Spring Harbor Laboratory Press; 2012.
- [34]Aziz R, Bartels D, Best A, DeJongh M, Disz T, Edwards R, Formsma K, Gerdes S, Glass E, Kubal M, Meyer F, Olsen G, Olson R, Osterman A, Overbeek R, McNeil L, Paarmann D, Paczian T, Parrello B, Pusch G, Reich C, Stevens R, Vassieva O, Vonstein V, Wilke A, Zagnitko O: The RAST server: rapid annotations using subsystems technology.BMC Genomics 2008, 9(1):75.
- [35]Darling ACE, Mau B, Blattner FR, Perna NT: Mauve: multiple alignment of conserved genomic sequence with rearrangements. Genome Res 2004, 14(7):1394-1403.
- [36]Hall BG: Building phylogenetic trees from molecular data with MEGA. Mol Biol Evol 2013, 30(5):1229-1235.
- [37]Saitou N, Nei M: The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 1987, 4(4):406-425.
- [38]Johnson JR, Stell AL: Extended virulence genotypes of Escherichia coli strains from patients with urosepsis in relation to phylogeny and host compromise. J Infect Dis 2000, 181(1):261-272.
- [39]Datsenko KA, Wanner BL: One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Proc Natl Acad Sci 2000, 97(12):6640-6645.
- [40]El-Mansi M, Anderson KJ, Inche CA, Knowles LK, Platt DJ: Isolation and curing of the Klebsiella pneumoniae large indigenous plasmid using sodium dodecyl sulphate. Res Microbiol 2000, 151(3):201-208.
- [41]Dudley EG, Abe C, Ghigo J-M, Latour-Lambert P, Hormazabal JC, Nataro JP: An IncI1 plasmid contributes to the adherence of the atypical enteroaggregative Escherichia coli Strain C1096 to cultured cells and abiotic surfaces. Infect Immun 2006, 74(4):2102-2114.