Recognition of the cellular beta1-chain integrin by the bacterial AfaD invasin is implicated in the internalization of afa-expressing pathogenic Escherichia coli strains.
The afa operons from Escherichia coli associated with extra-intestinal and intestinal infections have been characterized and the AfaD protein has been shown to be involved in the low internalization of laboratory strains expressing the afa-3 operon. The aim of this study was to determine the role of the AfaD invasin during the interaction of pathogenic E. coli with epithelial cells. We show that AfaD is implicated in the entry of a clinical isolate into both HeLa and undifferentiated Caco-2 cells. Once in the cytoplasm of these cells, the bacteria formed inclusions in which they were able to survive for at least 72 h. Internalization assays using polystyrene beads coated with His6-tagged purified AfaD (rAfaD) demonstrated that this invasin mediates entry into cells derived from various tissues (intestine and urothelium) that are targets for afa-positive strains. Consistent with the previous observation that an antibody blockade involving anti-alpha5beta1 integrin abolishes bacterial internalization, we show here that the entry of rAfaD-coated beads was dependent on the production and accessibility of beta1 integrins on the cells. The AfaD proteins belong to a family of invasins that are at least 45% identical. Despite their differences, the recombinant rAfaD-III and rAfaD-VIII proteins both bound to beta1 integrins. Our results suggest that beta1 integrin is a common receptor for AfaD invasins and that additional AfaD-type-specific receptors exist.
Plançon
Laure
L
Unité de Pathogénie Bactérienne des Muqueuses, Institut Pasteur 28 rue du Dr Roux, F-75724 Paris, France.
Du Merle
Laurence
L
Le Friec
Sandrine
S
Gounon
Pierre
P
Jouve
Mabel
M
Guignot
Julie
J
Servin
Alain
A
Le Bouguénec
Chantal
C
D018830:Adhesins, Escherichia coli / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D000818:Animals; D002460:Cell Line; D045744:Cell Line, Tumor; D015169:Colony Count, Microbial; D006224:Cricetinae; D004285:Dogs; D004847:Epithelial Cells / Q000166:cytology / Q000382:microbiology / Q000648:ultrastructure; D004926:Escherichia coli / Q000472:pathogenicity*; D004927:Escherichia coli Infections; D017353:Gene Deletion; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D006801:Humans; D007118:Immunoassay; D002479:Inclusion Bodies / Q000382:microbiology; D019012:Integrin beta1 / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D051379:Mice; D016254:Mutagenesis, Insertional / Q000235:genetics; D011994:Recombinant Proteins / Q000378:metabolism
D018830:Adhesins, Escherichia coli; C108510:AfaD protein, E coli; D019012:Integrin beta1; D011994:Recombinant Proteins