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| Content Provider | PubMed Central |
|---|---|
| Author | Cappello, Renato E. Guadalupe, Estrada-gutierrez Irles, Claudine Silvia, Giono-cerezo Bloch, Robert J. Nataro, James P. |
| Abstract | Enteroaggregative Escherichia coli (EAEC) is an emerging diarrheal pathogen. Many EAEC strains produce the plasmid encoded toxin (Pet), which elicits cytotoxic effects on human intestinal tissue. Pet-intoxicated HEp-2 cells exhibit rounding and detachment from the substratum, accompanied by loss of F-actin stress fibers and condensation of the spectrin-containing membrane cytoskeleton. Although studies suggest that Pet directly cleaves spectrin, it is not known if this is the essential mode of action of the toxin. In addition, the effects of Pet on cytoskeletal elements other than actin and spectrin have not been reported. Here, we demonstrate by immunofluorescence that upon Pet intoxication, HEp-2 and HT29 cells lose focal adhesion complexes (FAC), a process that includes redistribution of focal adhesion kinase (FAK), α-actinin, paxillin, vinculin, F-actin, and spectrin itself. This redistribution was coupled with depletion of phosphotyrosine labeling at FACs. Immunoblotting and immunoprecipitation experiments revealed that FAK was tyrosine dephosphorylated, prior to the redistribution of FAK and spectrin. Moreover, phosphatase inhibition blocked cell retraction, suggesting that tyrosine dephosphorylation is an event that precedes FAK cleavage. Finally, we show that in vitro tyrosine-dephophorylated FAK was susceptible to Pet cleavage. These data suggest that mechanisms other than spectrin redistribution occur during Pet intoxication. |
| Related Links | http://dx.doi.org/10.1111/j.1574-695x.2010.00776.x |
| Ending Page | 314 |
| Page Count | 14 |
| Starting Page | 301 |
| File Format | |
| ISSN | 09288244 |
| e-ISSN | 1574695X |
| Journal | FEMS immunology and medical microbiology |
| Issue Number | 3 |
| Volume Number | 61 |
| Language | English |
| Publisher Date | 2011-04-01 |
| Access Restriction | Open |
| Subject Keyword | Immunology Immunology and Allergy Microbiology (medical) Microbiology Infectious Diseases Research in Higher Education |
| Content Type | Text |
| Resource Type | Article |
| Subject | Infectious Diseases Immunology and Allergy Immunology Microbiology Microbiology (medical) |
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