Proteins other than the locus of enterocyte effacement-encoded proteins contribute to <it>Escherichia coli</it> O157:H7 adherence to bovine rectoanal junction stratified squamous epithelial cells
<p>Abstract</p> <p>Background</p> <p>In this study, we present evidence that proteins encoded by the Locus of Enterocyte Effacement (LEE), considered critical for <it>Escherichia coli</it> O157 (O157) adherence to follicle-associated epithelial (FAE) cells a...
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| Format: | Article |
| Language: | English |
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BMC
2012-06-01
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| Series: | BMC Microbiology |
| Subjects: | |
| Online Access: | http://www.biomedcentral.com/1471-2180/12/103 |
| _version_ | 1811311205089804288 |
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| author | Kudva Indira T Griffin Robert W Krastins Bryan Sarracino David A Calderwood Stephen B John Manohar |
| author_facet | Kudva Indira T Griffin Robert W Krastins Bryan Sarracino David A Calderwood Stephen B John Manohar |
| author_sort | Kudva Indira T |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>In this study, we present evidence that proteins encoded by the Locus of Enterocyte Effacement (LEE), considered critical for <it>Escherichia coli</it> O157 (O157) adherence to follicle-associated epithelial (FAE) cells at the bovine recto-anal junction (RAJ), do not appear to contribute to O157 adherence to squamous epithelial (RSE) cells also constituting this primary site of O157 colonization in cattle.</p> <p>Results</p> <p>Antisera targeting intimin-γ, the primary O157 adhesin, and other essential LEE proteins failed to block O157 adherence to RSE cells, when this pathogen was grown in DMEM, a culture medium that enhances expression of LEE proteins. In addition, RSE adherence of a DMEM-grown-O157 mutant lacking the intimin protein was comparable to that seen with its wild-type parent O157 strain grown in the same media. These adherence patterns were in complete contrast to that observed with HEp-2 cells (the adherence to which is mediated by intimin-γ), assayed under same conditions. This suggested that proteins other than intimin-γ that contribute to adherence to RSE cells are expressed by this pathogen during growth in DMEM. To identify such proteins, we defined the proteome of DMEM-grown-O157 (DMEM-proteome). GeLC-MS/MS revealed that the O157 DMEM-proteome comprised 684 proteins including several components of the cattle and human O157 immunome, orthologs of adhesins, hypothetical secreted and outer membrane proteins, in addition to the known virulence and LEE proteins. Bioinformatics-based analysis of the components of the O157 DMEM proteome revealed several new O157-specific proteins with adhesin potential.</p> <p>Conclusion</p> <p>Proteins other than LEE and intimin-γ proteins are involved in O157 adherence to RSE cells at the bovine RAJ. Such proteins, with adhesin potential, are expressed by this human pathogen during growth in DMEM. Ongoing experiments to evaluate their role in RSE adherence should provide both valuable insights into the O157-RSE interactions and new targets for more efficacious anti-adhesion O157 vaccines.</p> |
| first_indexed | 2024-04-13T10:13:39Z |
| format | Article |
| id | doaj.art-fea45428a7a44ffea828f472205c9a23 |
| institution | Directory Open Access Journal |
| issn | 1471-2180 |
| language | English |
| last_indexed | 2024-04-13T10:13:39Z |
| publishDate | 2012-06-01 |
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| series | BMC Microbiology |
| spelling | doaj.art-fea45428a7a44ffea828f472205c9a232022-12-22T02:50:49ZengBMCBMC Microbiology1471-21802012-06-0112110310.1186/1471-2180-12-103Proteins other than the locus of enterocyte effacement-encoded proteins contribute to <it>Escherichia coli</it> O157:H7 adherence to bovine rectoanal junction stratified squamous epithelial cellsKudva Indira TGriffin Robert WKrastins BryanSarracino David ACalderwood Stephen BJohn Manohar<p>Abstract</p> <p>Background</p> <p>In this study, we present evidence that proteins encoded by the Locus of Enterocyte Effacement (LEE), considered critical for <it>Escherichia coli</it> O157 (O157) adherence to follicle-associated epithelial (FAE) cells at the bovine recto-anal junction (RAJ), do not appear to contribute to O157 adherence to squamous epithelial (RSE) cells also constituting this primary site of O157 colonization in cattle.</p> <p>Results</p> <p>Antisera targeting intimin-γ, the primary O157 adhesin, and other essential LEE proteins failed to block O157 adherence to RSE cells, when this pathogen was grown in DMEM, a culture medium that enhances expression of LEE proteins. In addition, RSE adherence of a DMEM-grown-O157 mutant lacking the intimin protein was comparable to that seen with its wild-type parent O157 strain grown in the same media. These adherence patterns were in complete contrast to that observed with HEp-2 cells (the adherence to which is mediated by intimin-γ), assayed under same conditions. This suggested that proteins other than intimin-γ that contribute to adherence to RSE cells are expressed by this pathogen during growth in DMEM. To identify such proteins, we defined the proteome of DMEM-grown-O157 (DMEM-proteome). GeLC-MS/MS revealed that the O157 DMEM-proteome comprised 684 proteins including several components of the cattle and human O157 immunome, orthologs of adhesins, hypothetical secreted and outer membrane proteins, in addition to the known virulence and LEE proteins. Bioinformatics-based analysis of the components of the O157 DMEM proteome revealed several new O157-specific proteins with adhesin potential.</p> <p>Conclusion</p> <p>Proteins other than LEE and intimin-γ proteins are involved in O157 adherence to RSE cells at the bovine RAJ. Such proteins, with adhesin potential, are expressed by this human pathogen during growth in DMEM. Ongoing experiments to evaluate their role in RSE adherence should provide both valuable insights into the O157-RSE interactions and new targets for more efficacious anti-adhesion O157 vaccines.</p>http://www.biomedcentral.com/1471-2180/12/103O157Rectoanal junctionLEEAdherenceDMEMGeLC-MS/MS |
| spellingShingle | Kudva Indira T Griffin Robert W Krastins Bryan Sarracino David A Calderwood Stephen B John Manohar Proteins other than the locus of enterocyte effacement-encoded proteins contribute to <it>Escherichia coli</it> O157:H7 adherence to bovine rectoanal junction stratified squamous epithelial cells BMC Microbiology O157 Rectoanal junction LEE Adherence DMEM GeLC-MS/MS |
| title | Proteins other than the locus of enterocyte effacement-encoded proteins contribute to <it>Escherichia coli</it> O157:H7 adherence to bovine rectoanal junction stratified squamous epithelial cells |
| title_full | Proteins other than the locus of enterocyte effacement-encoded proteins contribute to <it>Escherichia coli</it> O157:H7 adherence to bovine rectoanal junction stratified squamous epithelial cells |
| title_fullStr | Proteins other than the locus of enterocyte effacement-encoded proteins contribute to <it>Escherichia coli</it> O157:H7 adherence to bovine rectoanal junction stratified squamous epithelial cells |
| title_full_unstemmed | Proteins other than the locus of enterocyte effacement-encoded proteins contribute to <it>Escherichia coli</it> O157:H7 adherence to bovine rectoanal junction stratified squamous epithelial cells |
| title_short | Proteins other than the locus of enterocyte effacement-encoded proteins contribute to <it>Escherichia coli</it> O157:H7 adherence to bovine rectoanal junction stratified squamous epithelial cells |
| title_sort | proteins other than the locus of enterocyte effacement encoded proteins contribute to it escherichia coli it o157 h7 adherence to bovine rectoanal junction stratified squamous epithelial cells |
| topic | O157 Rectoanal junction LEE Adherence DMEM GeLC-MS/MS |
| url | http://www.biomedcentral.com/1471-2180/12/103 |
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