Bacterial secreted proteins are required for the internaliztion of Campylobacter jejuni into cultured mammalian cells.
Presented here is the first evidence that Campylobacter jejuni secrete proteins upon co-cultivation with host cells and in INT 407 cell-conditioned medium. A C. jejuni gene designated ciaB for Campylobacter invasion antigen B was identified, using a differential screening technique, which is required for this secretion process and the efficient entry of this bacterium into a host cell. The C. jejuni ciaB gene encodes a protein of 610 amino acids with a calculated molecular mass of 73 154 Da. The deduced amino acid sequence of the CiaB protein shares similarity with type III secreted proteins associated with the invasion of host cells from other more extensively characterized bacterial pathogens. In vitro binding and internalization assays revealed that the binding of C. jejuni ciaB null mutants was indistinguishable from that of the parental isolate, whereas a significant reduction was noted in internalization. Confocal microscopic examination of C. jejuni-infected cells revealed that CiaB was translocated into the cytoplasm of the host cells. Culturing C. jejuni with INT 407 cells or in INT 407-conditioned medium resulted in the secretion of at least eight proteins, ranging in size from 12.8 to 108 kDa, into the culture medium. C. jejuni ciaB null mutants were deficient in the secretion of all eight proteins, indicating that CiaB is required for the secretion process. The identification of the C. jejuni ciaB gene represents a significant advance in understanding the molecular mechanism of C. jejuni internalization and the pathogenesis of C. jejuni-mediated enteritis.
Konkel
M E
ME
Department of Microbiology, Washington State University, Pullman, Washington 99164-4233, USA. Konkel@mail.wsu.edu
Kim
B J
BJ
Rivera-Amill
V
V
Garvis
S G
SG
D000595:Amino Acid Sequence; D000942:Antigens, Bacterial / Q000737:chemistry / Q000235:genetics*; D016123:Campylobacter jejuni / Q000235:genetics* / Q000472:pathogenicity; D002460:Cell Line; D003001:Cloning, Molecular; D018920:Coculture Techniques; D017077:Culture Media, Conditioned; D015723:Gene Library; D005798:Genes, Bacterial; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D010957:Plasmids; D011994:Recombinant Proteins / Q000235:genetics; D016415:Sequence Alignment; D015398:Signal Transduction
D000942:Antigens, Bacterial; C120454:CiaB protein, Campylobacter jejuni; D017077:Culture Media, Conditioned; D011994:Recombinant Proteins