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研究生(外文):Nien-Pei Tsai
論文名稱(外文):Multiple Functions of L0036 in Regulation of the Pathogenicity Island of Enterohemorrhagic Escherichia coli O157:H7
指導教授(外文):Wan-Jr Syu
外文關鍵詞:Enterohemorrhagic Escherichia coli O157:H7regulationLEE island
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出血性大腸桿菌與致病性大腸桿菌 (EPEC) 都屬於病原性的大腸桿菌,其感染人類腸內上皮細胞後會造成腸內微絨毛的破壞,引發嚴重的所謂 A/E lesion 病灶。一般來說,引發 A/E lesion 病原基因均來自於細菌基因組中一段被稱為 LEE island 的特殊區域,此區域共包含有 41 個開放閱讀框架 (open reading frame),而約可歸為五個主要的操縱子 (operon),分別為 LEE1、LEE2、LEE3、LEE4 與 tir (LEE5)。LEE island 的表現受到非常複雜的調控,就目前所知,在 LEE island 有兩個活化型調控蛋白 (Ler 與 GrlA) 與一個抑制型調控蛋白 (GrlR) 的存在 (Deng et al.),然而我們將在這裡引入一個尚未發表的調控性蛋白質 L0036。在已發表的文獻中,L0036 曾被推斷為形成第三型分泌系統的一個組成分子(Aizawa, 2001),不過在此我們發現 L0036 是個在 RNA 與 蛋白質層面的表現都受到嚴謹調控的基因,但 L0036 的表現與否並不影響第三型分泌系統的正常功能,然而 L0036 下游基因的轉譯卻又仰賴 L0036 的完整轉譯,可見其重要性;另外,當我們將 L0036 過量表現於 EHEC 中時,我們發現 L0036 會專一性的與 Ler 產生交互作用進而抑制 LEE island 的表現。由於 L0036 具有此多重功能,因此我們將 L0036 重新命名為 multiple-point controller (mpc) 而其基因產物則為 Mpc。
Diarrheagenic enterohemorrhagic Escherichia coli (EHEC) and enteropathogenic E. coli (EPEC) attach to human intestinal epithelium and efface brush border microvilli, forming A/E lesions. These human pathogens are phenotypically similar to the mouse pathogen Citrobacter rodentium. Genetically, they all have a homologous set of virulent genes involved in the A/E lesion, and these genes are organized on a locus of enterocyte effacement (Deng et al.), a pathogenicity island. The LEE island comprises 41 specific open reading frames (orfs), of which most are organized in five operons, LEE1, LEE2, LEE3, LEE4, and tir (LEE5). The expression of the LEE genes is regulated in a complicated manner, and current knowledge is that there are at least two positive regulators, Ler and GrlA, and one negative regulator, called GrlR. In EHEC, GrlA is encoded by L0043 whereas GrlR is encoded by L0044. In this thesis, we report a third regulatory gene located in LEE3, i.e., orf L0036 in EHEC. In the previous studies, L0036 has been implicated to function as a component of TTSS since the corresponding mutant phenotypically behaved as those with deletion of other orfs on the same operon or on LEE2. In addition to this observation, we found its expression was tightly controlled at both mRNA and protein levels. When overexpressed, this factor, named Mpc, interacted with Ler and suppressed the expression of the LEE proteins. When the translation wass not initiated or terminated before maturation, the type III secretion of effectors was completely abolished. Therefore, L0036 appeared to have multiple functions in addition to the fact that several cis elements reside on the region where L0036 spans.
Abe, A., de Grado, M., Pfuetzner, R.A., Sanchez-Sanmartin, C., Devinney, R., Puente,
J.L., Strynadka, N.C., and Finlay, B.B. (1999) Enteropathogenic Escherichia coli
translocated intimin receptor, Tir, requires a specific chaperone for stable secretion.
Mol Microbiol 33: 1162-1175.
Aiba, H., Adhya, S., and de Crombrugghe, B. (1981) Evidence for two functional gal
promoters in intact Escherichia coli cells. J Biol Chem 256: 11905-11910.
Aizawa, S.I. (2001) Bacterial flagella and type III secretion systems. FEMS Microbiol
Lett 202: 157-164.
Bustamante, V.H., Santana, F.J., Calva, E., and Puente, J.L. (2001) Transcriptional
regulation of type III secretion genes in enteropathogenic Escherichia coli: Ler
antagonizes H-NS-dependent repression. Mol Microbiol 39: 664-678.
Creasey, E.A., Delahay, R.M., Bishop, A.A., Shaw, R.K., Kenny, B., Knutton, S., and
Frankel, G. (2003a) CesT is a bivalent enteropathogenic Escherichia coli chaperone
required for translocation of both Tir and Map. Mol Microbiol 47: 209-221.
Creasey, E.A., Friedberg, D., Shaw, R.K., Umanski, T., Knutton, S., Rosenshine, I.,
and Frankel, G. (2003b) CesAB is an enteropathogenic Escherichia coli chaperone for
the type-III translocator proteins EspA and EspB. Microbiology 149: 3639-3647.
Deng, W., Puente, J.L., Gruenheid, S., Li, Y., Vallance, B.A., Vazquez, A., Barba, J.,
Ibarra, J.A., O''Donnell, P., Metalnikov, P., Ashman, K., Lee, S., Goode, D., Pawson,
T., and Finlay, B.B. (2004) Dissecting virulence: systematic and functional analyses
of a pathogenicity island. Proc Natl Acad Sci U S A 101: 3597-3602.
Elliott, S.J., Wainwright, L.A., McDaniel, T.K., Jarvis, K.G., Deng, Y.K., Lai, L.C.,
McNamara, B.P., Donnenberg, M.S., and Kaper, J.B. (1998) The complete sequence
of the locus of enterocyte effacement (LEE) from enteropathogenic Escherichia coli
E2348/69. Mol Microbiol 28: 1-4.
Elliott, S.J., Sperandio, V., Giron, J.A., Shin, S., Mellies, J.L., Wainwright, L.,
Hutcheson, S.W., McDaniel, T.K., and Kaper, J.B. (2000) The locus of enterocyte
effacement (LEE)-encoded regulator controls expression of both LEE- and
non-LEE-encoded virulence factors in enteropathogenic and enterohemorrhagic
Escherichia coli. Infect Immun 68: 6115-6126.
Elliott, S.J., Krejany, E.O., Mellies, J.L., Robins-Browne, R.M., Sasakawa, C., and
Kaper, J.B. (2001) EspG, a novel type III system-secreted protein from
enteropathogenic Escherichia coli with similarities to VirA of Shigella flexneri. Infect
Immun 69: 4027-4033.
Friedberg, D., Umanski, T., Fang, Y., and Rosenshine, I. (1999) Hierarchy in the
expression of the locus of enterocyte effacement genes of enteropathogenic
Escherichia coli. Mol Microbiol 34: 941-952.
Gauthier, A., Puente, J.L., and Finlay, B.B. (2003) Secretin of the enteropathogenic
Escherichia coli type III secretion system requires components of the type III
apparatus for assembly and localization. Infect Immun 71: 3310-3319.
Gomez-Duarte, O.G., and Kaper, J.B. (1995) A plasmid-encoded regulatory region
activates chromosomal eaeA expression in enteropathogenic Escherichia coli. Infect
Immun 63: 1767-1776.
Griffin, P.M., and Tauxe, R.V. (1991) The epidemiology of infections caused by
Escherichia coli O157:H7, other enterohemorrhagic E. coli, and the associated
hemolytic uremic syndrome. Epidemiol Rev 13: 60-98.
Haack, K.R., Robinson, C.L., Miller, K.J., Fowlkes, J.W., and Mellies, J.L. (2003)
Interaction of Ler at the LEE5 (tir) operon of enteropathogenic Escherichia coli. Infect
Immun 71: 384-392.
Ide, T., Laarmann, S., Greune, L., Schillers, H., Oberleithner, H., and Schmidt, M.A.
(2001) Characterization of translocation pores inserted into plasma membranes by
type III-secreted Esp proteins of enteropathogenic Escherichia coli. Cell Microbiol 3:
Jarvis, K.G., Giron, J.A., Jerse, A.E., McDaniel, T.K., Donnenberg, M.S., and Kaper,
J.B. (1995) Enteropathogenic Escherichia coli contains a putative type III secretion
system necessary for the export of proteins involved in attaching and effacing lesion
formation. Proc Natl Acad Sci U S A 92: 7996-8000.
Jerse, A.E., and Kaper, J.B. (1991) The eae gene of enteropathogenic Escherichia coli
encodes a 94-kilodalton membrane protein, the expression of which is influenced by
the EAF plasmid. Infect Immun 59: 4302-4309.
Kenny, B., Lai, L.C., Finlay, B.B., and Donnenberg, M.S. (1996) EspA, a protein
secreted by enteropathogenic Escherichia coli, is required to induce signals in
epithelial cells. Mol Microbiol 20: 313-323.
Kenny, B., DeVinney, R., Stein, M., Reinscheid, D.J., Frey, E.A., and Finlay, B.B.
(1997) Enteropathogenic E. coli (EPEC) transfers its receptor for intimate adherence
into mammalian cells. Cell 91: 511-520.
Kenny, B., and Jepson, M. (2000) Targeting of an enteropathogenic Escherichia coli
(EPEC) effector protein to host mitochondria. Cell Microbiol 2: 579-590.
Kimbrough, T.G., and Miller, S.I. (2002) Assembly of the type III secretion needle
complex of Salmonella typhimurium. Microbes Infect 4: 75-82.
Knutton, S., Baldwin, T., Williams, P.H., and McNeish, A.S. (1989) Actin
accumulation at sites of bacterial adhesion to tissue culture cells: basis of a new
diagnostic test for enteropathogenic and enterohemorrhagic Escherichia coli. Infect
Immun 57: 1290-1298.
Koutkia, P., Mylonakis, E., and Flanigan, T. (1997) Enterohemorrhagic Escherichia
coli O157:H7--an emerging pathogen. Am Fam Physician 56: 853-856, 859-861.
Lai, L.C., Wainwright, L.A., Stone, K.D., and Donnenberg, M.S. (1997) A third
secreted protein that is encoded by the enteropathogenic Escherichia coli
pathogenicity island is required for transduction of signals and for attaching and
effacing activities in host cells. Infect Immun 65: 2211-2217.
Levine, M.M., Xu, J.G., Kaper, J.B., Lior, H., Prado, V., Tall, B., Nataro, J., Karch, H.,
and Wachsmuth, K. (1987) A DNA probe to identify enterohemorrhagic Escherichia
coli of O157:H7 and other serotypes that cause hemorrhagic colitis and hemolytic
uremic syndrome. J Infect Dis 156: 175-182.
Link, A.J., Phillips, D., and Church, G.M. (1997) Methods for generating precise
deletions and insertions in the genome of wild-type Escherichia coli: application to
open reading frame characterization. J Bacteriol 179: 6228-6237.
Mellies, J.L., Elliott, S.J., Sperandio, V., Donnenberg, M.S., and Kaper, J.B. (1999)
The Per regulon of enteropathogenic Escherichia coli : identification of a regulatory
cascade and a novel transcriptional activator, the locus of enterocyte effacement
(LEE)-encoded regulator (Ler). Mol Microbiol 33: 296-306.
Moon, H.W., Whipp, S.C., Argenzio, R.A., Levine, M.M., and Giannella, R.A. (1983)
Attaching and effacing activities of rabbit and human enteropathogenic Escherichia
coli in pig and rabbit intestines. Infect Immun 41: 1340-1351.
Mundy, R., Petrovska, L., Smollett, K., Simpson, N., Wilson, R.K., Yu, J., Tu, X.,
Rosenshine, I., Clare, S., Dougan, G., and Frankel, G. (2004) Identification of a novel
Citrobacter rodentium type III secreted protein, EspI, and roles of this and
other secreted proteins in infection. Infect Immun 72: 2288-2302.
Neves, B.C., Mundy, R., Petrovska, L., Dougan, G., Knutton, S., and Frankel, G.
(2003) CesD2 of enteropathogenic Escherichia coli is a second chaperone for the type
III secretion translocator protein EspD. Infect Immun 71: 2130-2141.
O''Brien, A.D., Lively, T.A., Chang, T.W., and Gorbach, S.L. (1983) Purification of
Shigella dysenteriae 1 (Shiga)-like toxin from Escherichia coli O157:H7 strain
associated with haemorrhagic colitis. Lancet 2: 573.
Perna, N.T., Mayhew, G.F., Posfai, G., Elliott, S., Donnenberg, M.S., Kaper, J.B., and
Blattner, F.R. (1998) Molecular evolution of a pathogenicity island from
enterohemorrhagic Escherichia coli O157:H7. Infect Immun 66: 3810-3817.
Riley, L.W. (1987) The epidemiologic, clinical, and microbiologic features of
hemorrhagic colitis. Annu Rev Microbiol 41: 383-407.
Romeo, T. (1998) Global regulation by the small RNA-binding protein CsrA and the
non-coding RNA molecule CsrB. Mol Microbiol 29: 1321-1330.
Schmidt, H., Karch, H., and Beutin, L. (1994) The large-sized plasmids of
enterohemorrhagic Escherichia coli O157 strains encode hemolysins which are
presumably members of the E. coli alpha-hemolysin family. FEMS Microbiol Lett
117: 189-196.
Schmidt, H., Beutin, L., and Karch, H. (1995) Molecular analysis of the
plasmid-encoded hemolysin of Escherichia coli O157:H7 strain EDL 933. Infect
Immun 63: 1055-1061.
Sperandio, V., Mellies, J.L., Nguyen, W., Shin, S., and Kaper, J.B. (1999) Quorum
sensing controls expression of the type III secretion gene transcription and protein
secretion in enterohemorrhagic and enteropathogenic Escherichia coli. Proc Natl Acad
Sci U S A 96: 15196-15201.
Sperandio, V., Mellies, J.L., Delahay, R.M., Frankel, G., Crawford, J.A., Nguyen, W.,
and Kaper, J.B. (2000) Activation of enteropathogenic Escherichia coli (EPEC) LEE2
and LEE3 operons by Ler. Mol Microbiol 38: 781-793.
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. J Bacteriol 178: 6546-6554.
Sukhan, A., Kubori, T., Wilson, J., and Galan, J.E. (2001) Genetic analysis of
assembly of the Salmonella enterica serovar Typhimurium type III
secretion-associated needle complex. J Bacteriol 183: 1159-1167.
Tu, X., Nisan, I., Yona, C., Hanski, E., and Rosenshine, I. (2003) EspH, a new
cytoskeleton-modulating effector of enterohaemorrhagic and enteropathogenic
Escherichia coli. Mol Microbiol 47: 595-606.
Wainwright, L.A., and Kaper, J.B. (1998) EspB and EspD require a specific
chaperone for proper secretion from enteropathogenic Escherichia coli. Mol Microbiol
27: 1247-1260.
Wells, J.G., Davis, B.R., Wachsmuth, I.K., Riley, L.W., Remis, R.S., Sokolow, R., and
Morris, G.K. (1983) Laboratory investigation of hemorrhagic colitis outbreaks
associated with a rare Escherichia coli serotype. J Clin Microbiol 18: 512-520.
Wilson, R.K., Shaw, R.K., Daniell, S., Knutton, S., and Frankel, G. (2001) Role of
EscF, a putative needle complex protein, in the type III protein translocation system of
enteropathogenic Escherichia coli. Cell Microbiol 3: 753-762.
吳宜之 (2002) 從訊息核醣核酸及蛋白質層面分析出血性大腸桿菌O157:H7 致病島嶼,碩士論文,國立陽明大學微生物及免疫學研究所。
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