ABC  Vol.2 No.3 , August 2012
Identification of the autotransporter Pet toxin in Proteus mirabilis strain isolated from patients with urinary tract infections
Abstract: Proteus mirabilis, a motile Gram-negative bacterium, represents a common cause of complicated urinary tract infections. Autotransporters are a family of secreted proteins from Gram-negative bacteria that direct their own secretion across the outer membrane (type V autotransporter secretion mechanism). Serine protease autotransporters of Enterobacteriaceae (SPATEs) include adhesins, toxins, and proteases that can contribute to the virulence. Plasmid-encoded toxin (Pet) is the predominant protein in culture supernatants of enteroaggregative E. coli prototype strain 042 and has been extensively studied. Pet toxin is encoded on the 65-MDa adherence-related plasmid of EAEC 042 strain. In this work, Pet protein was found in the supernatant obtained from Proteus mirabilis RTX339 strain isolated from a psychiatric patient suffering complicated urinary tract infections (UTIs). The nucleotide sequence of pet gene was obtained using primers designed from E. coli 042 pet gene reported. The alignment of the sequence showed 100% identity with the pet gene reported. Is important to note that Proteus mirabilis RTX339 pet gene has chromosomal location. The chromosomal location of the gene was established since no plasmids were harbored by this strain.
Cite this paper: Gutiérrez-Lucas, L. , Mendoza-Hernández, G. , González-Pedrajo, B. , Eslava-Campos, C. , Bustos-Martínez, J. and Sainz-Espuñes, T. (2012) Identification of the autotransporter Pet toxin in Proteus mirabilis strain isolated from patients with urinary tract infections. Advances in Biological Chemistry, 2, 283-290. doi: 10.4236/abc.2012.23036.

[1]   Chippendale, G.R., Warren, J.W., Trifillis, A.L. and Mobley, H.L.T. (1994) Internalization of Proteus mirabilis by human renal epithelial cells. Infection and Immunity, 62, 3115-3121.

[2]   Allison, C., Coleman, N., Jones, P.L. and Hughes, C. (1992) Ability of Proteus mirabilis to invade human urothelial cells is coupled to motility and swarming differentiation. Infection and Immunity, 60, 4740-4746.

[3]   Jiang, S., Lin, T., Wang, W., Liu, M., Hsueh, P. and Liaw, S. (2010) Characterization of UDP-Glucose dehydrogenase and UDP-Glucose pyrophosphorylase mutants of Proteus mirabilis: Defectiveness in polymyxin B resistance, swarming, and virulence. Antimicrobial Agents and Chemotherapy, 54, 2000-2009. doi:10.1128/AAC.01384-09

[4]   Stickler, D.J., Jones, S.M., Adusei, G.O. and Waters, M.G. (2006) A sensor to detect the early stages in the development of crystalline Proteus mirabilis biofilm on indwelling bladder catheters. Journal of Clinical Microbiology, 38, 1540-1542. doi:10.1128/JCM.44.4.1540-1542.2006

[5]   Jacobsen, S.M., Stickler, D.J., Mobley, H.L.T. and Shirtliff, M.E. (2008) Complicated catheter-associated urinary tract infections due to Escherichia coli and Proteus mirabilis. Clinical Microbiology Reviews, 21, 26-59. doi:10.1128/CMR.00019-07

[6]   Restieri. C., Garriss, G., Locas, M. and Dozois, C. (2007) Autotransporter-encoding sequences are phylogenetically distributed among Escherichia coli clinical isolates and reference strains. Applied and Environmental Microbiology, 73, 1553-1562. doi:10.1128/AEM.01542-06

[7]   Eslava, C., Navarro-García, F., Czeczulin, J., Henderson, I., Cravioto, A., et al. (1998) Pet, an autotransporter enterotoxin from enteroaggregative Escherichia coli. Infection and Immunity, 66, 3155-3163.

[8]   Navarro-García, F., Canizales-Roman, A., Burlingame, K., Teter, K. and Vidal, J. (2007) Pet, a non-AB toxin, is transported and translocated into epithelial cells by a retrograde trafficking pathway. Infection and Immunity, 75, 2101-2109. doi:10.1128/IAI.01515-06

[9]   Villaseca, J., Navarro-García, F., Mendoza-Hernández, G., Nataro, J., Cravioto, A., et al. (2000) Pet toxin from enteroaggregative Escherichia coli produces cellular damage associated with fodrin disruption. Infection and Immunity, 68, 5920-5927. doi:10.1128/IAI.68.10.5920-5927.2000

[10]   Sainz, T., Perez, J., Fresan, Ma.C., Flores, V., Jimenez, L., et al. (2002) Histological alterations and immune response induced by Pet toxin during colonization with Enteroaggregative Escherichia coli (EAEC) in a mouse model infection. Korean Journal of Microbiology, 40, 91- 97.

[11]   Pearson, M., Sebaihia, M., Churcher, C., Quail, M., Seshasayee, A., et al. (2008) Complete genome sequence of uropathogenic Proteus mirabilis, a master of both adherence and motility. Journal of Bacteriology, 190, 4027- 4037. doi:10.1128/JB.01981-07

[12]   Alamuri, P. and Mobley, H.L.T. (2008) A novel autotransporter of uropathogenic Proteus mirabilis is both a cytotoxin and an agglutinin. Molecular Microbiology, 68, 997-1017. doi:10.1111/j.1365-2958.2008.06199.x

[13]   Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227, 680-685. doi:10.1038/227680a0

[14]   Towbin, H., Staehelin, T. and Gordon, J. (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. Proceedings of the National Academy of Sciences USA, 76, 4350-4354. doi:10.1073/pnas.76.9.4350

[15]   Bienvenut, W.V., Sanchez, J.C., Karmime, A., Rougue, V., Rose, K., et al. (1999) Toward a clinical molecular scanner for proteome research: Parallel protein chemical processing before and during western blot. Analytical Chemistry, 21, 4800. doi:10.1021/ac990448m

[16]   Kinter, M., Sherman, N.E., Desiderio, D.M. and Nibbering, N.M.M. (2000) In protein sequencing and identification using tandem mass spectrometry. John Wiley-Inter-science, Inc., New York, 147-165.

[17]   Bourbeau, P. and Heiter, B. (1998) Comparison of Vitek GNI and GNI+ Cards for Identification of Gram-Negative Bacteria. Journal of Clinical Microbiology, 36, 2775- 2777.

[18]   Nielubowicz, G., Smith, S. and Mobley, H.L.T. (2010) Zinc uptake contributes to motility and provides a competitive advantage to Proteus mirabilis during experimental urinary tract infection. Infection and Immunity, 78, 2823-2833. doi:10.1128/IAI.01220-09

[19]   Jansen, A., Lockatell, C., Johnson, D. and Mobley, H.L.T. (2003) Visualization of Proteus mirabilis morphotypes in the urinary tract: the elongated swarmer cell is rarely observed in ascending urinary tract infection. Infection and Immunity, 71, 3607-3613. doi:10.1128/IAI.71.6.3607-3613.2003

[20]   Guyer, D., Kao, J. and Mobley, H.L.T. (1998) Genomic analysis of a pathogenicity island in uropathogenic Escherichia coli CFT073: Distribution of homologous sequences among isolates from patients with pyelonephritis, cystitis, and catheter-associated bacteriuria and from fecal samples. Infection and Immunity, 66, 4411-4417.

[21]   Dutta, P., Sui, B., Nataro, J. (2003) Structure-function analysis of the enteroaggregative Escherichia coli plasmid-encoded toxin autotransporter using scanning linker mutagenesis. Journal of Biological Chemistry, 278, 39912-39920. doi:10.1074/jbc.M303595200

[22]   Henderson, I. R., Czeczulin, J., Eslava, C., Noriega, F. and Nataro, J. (1999) Characterization of Pic, a secreted protease of Shigella flexneri and enteroaggregative Escherichia coli. Infection and Immunity, 67, 5587-5596.

[23]   Henderson, I. and Nataro, J. (2001) Virulence functions of autotransporter proteins. Infection and Immunity, 69, 1231-1243. doi:10.1128/IAI.69.3.1231-1243.2001

[24]   Sui, B., Dutta, P., Nataro, J. (2003) Intracellular expression of the plasmid-encoded toxin from enteroaggregative Escherichia coli. Infection and Immunity, 71, 5364-5370. doi:10.1128/IAI.71.9.5364-5370.2003

[25]   Al-Hasani, K., Henderson, I., Sakellaris, H., Rajakumar, K., Grant, T., et al. (2000) The sigA gene which is borne on the she pathogenicity island of Shigella flexneri 2a encodes an exported cytopathic protease involved in intestinal fluid accumulation. Infection and Immunity, 68, 2457-2463. doi:10.1128/IAI.68.5.2457-2463.2000

[26]   Benjelloun-Touimi, Z., Sansonetti, P. and Parsot, C. (1995) SepA, the major extracellular protein of Shigella flexneri: autonomous secretion and involvement in tissue invasion. Molecular Microbiology, 17, 123-135. doi:10.1111/j.1365-2958.1995.mmi_17010123.x

[27]   Guyer, D. M., Henderson, I. R., Nataro, J. P., Mobley and H. L. (2000) Identification of sat, an autotransporter toxin produced by uropathogenic Escherichia coli. Molecular Microbiology, 38, 53-66. doi:10.1046/j.1365-2958.2000.02110.x

[28]   Brunder, W., Schmidt, H., Karch, H. (1997) EspP, a novel extracellular serine protease of enterohaemorrhagic Escherichia coli O157:H7 cleaves human coagulation factor V. Molecular Microbiology, 24, 767-778. doi:10.1046/j.1365-2958.1997.3871751.x

[29]   Mellies, J.L., Navarro-García, F., Okeke, I., Frederickson, J., Nataro, J.P., et al. (2001) EspC pathogenicity island of enteropathogenic Escherichia coli encodes an enterotoxin. Infection and Immunity, 69, 315-324. doi:10.1128/IAI.69.1.315-324.2001

[30]   Stein, M., Kenny, B., Stein, M.A. and Finlay, B.B. (1996) Characterization of EspC, a 110-kilodalton protein secreted by enteropathogenic Escherichia coli which is homologous to members of the immunoglobulin A protease-like family of secreted proteins. Journal of Bacteriology, 178, 6546-6554.

[31]   Behrens, M., Sheikh, J. and Nataro, J. P. (2002) Regulation of the overlapping pic/set locus in Shigella flexneri and enteroaggregative Escherichia coli. Infection and Immunity, 70, 2915-2925. doi:10.1128/IAI.70.6.2915-2925.2002