Initial binding of Shiga toxin-producing Escherichia coli to host cells and subsequent induction of actin rearrangements depend on filamentous EspA-containing surface appendages.
Shiga toxin-producing Escherichia coli (STEC) induce so-called attaching and effacing lesions that enable the tight adherence of these pathogens to the gut epithelium. All of the genes necessary for this process are present in the locus of enterocyte effacement, which encodes a type III secretion system, the secreted Esp proteins and the surface protein intimin. In this study we sequenced the espA gene of STEC, generated and characterized a corresponding deletion mutant and raised EspA-specific monoclonal antibodies to analyse the functional role of this protein during infection. EspA was detected in often filament-like structures decorating all bacteria that had attached to HeLa cells. These appendages were especially prominent on bacteria that had not yet induced the formation of actin pedestals, indicating that they mediate the initial contact of STEC to their target cells. Consistently, a deletion of the espA gene completely abolished the capacity of such STEC mutants to bind to HeLa cells and to induce actin rearrangements. Surface appendages similar to those described in this study are also formed by Pseudomonas syringae and may represent a structural element common to many bacterial pathogens that deliver proteins into their target cells via a type III secretion system.
Ebel
F
F
Institut für Medizinische Mikrobiologie, Justus-Liebig-Universität, Giessen, Germany. frank.ebel@mikrobio.med.uni-giessen
Podzadel
T
T
Rohde
M
M
Kresse
A U
AU
Krämer
S
S
Deibel
C
C
Guzmán
C A
CA
Chakraborty
T
T
D000199:Actins / Q000378:metabolism*; D018829:Adhesins, Bacterial; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D001422:Bacterial Adhesion; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D002352:Carrier Proteins; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development / Q000472:pathogenicity*; D019453:Escherichia coli O157 / Q000235:genetics / Q000472:pathogenicity; D029968:Escherichia coli Proteins; D005408:Flagellin / Q000235:genetics; D017353:Gene Deletion; D006367:HeLa Cells; D006801:Humans; D008264:Macrophages / Q000382:microbiology; D051379:Mice; D018613:Microscopy, Confocal; D008855:Microscopy, Electron, Scanning; D016253:Microscopy, Immunoelectron; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA; D015398:Signal Transduction
D000199:Actins; D018829:Adhesins, Bacterial; D000911:Antibodies, Monoclonal; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D002352:Carrier Proteins; D029968:Escherichia coli Proteins; C102051:EspA protein, E coli; D005408:Flagellin; C073819:eaeA protein, E coli