Interactions of Campylobacter jejuni cytolethal distending toxin subunits CdtA and CdtC with HeLa cells.
Campylobacter jejuni produces a toxin, called cytolethal distending toxin (CDT), which causes direct DNA damage leading to invocation of DNA damage checkpoint pathways. The affected cells arrest in G(1) or G(2) and eventually die. CDT consists of three protein subunits, CdtA, CdtB, and CdtC, with CdtB recently identified as a nuclease. However, little is known about the functions of CdtA or CdtC. In this work, enzyme-linked immunosorbent assay-based experiments were used to show, for the first time, that both CdtA and CdtC bound with specificity to the surface of HeLa cells, whereas CdtB did not. Varying the order of the addition of subunits for reconstitution of the holotoxin had no effect on activity. In addition, mutants containing deletions of conserved regions of CdtA and CdtC were able to bind to the surface of HeLa cells but were not able to participate in holotoxin assembly. Finally, both Cdt mutant subunits were able to effectively compete with CDT holotoxin in the HeLa cell binding assay.
Lee
Robert B
RB
Department of Microbiology, Immunology, and Molecular Genetics, School of Medicine, University of Kentucky, Lexington, Kentucky 40536, USA.
Hassane
Duane C
DC
Cottle
Daniel L
DL
Pickett
Carol L
CL
D000595:Amino Acid Sequence; D001427:Bacterial Toxins / Q000737:chemistry / Q000235:genetics / Q000633:toxicity*; D001483:Base Sequence; D001667:Binding, Competitive; D001710:Biotin; D016123:Campylobacter jejuni / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity* / Q000502:physiology; D004269:DNA, Bacterial / Q000235:genetics; D005798:Genes, Bacterial; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D011485:Protein Binding; D021122:Protein Subunits; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000633:toxicity
D001427:Bacterial Toxins; D004269:DNA, Bacterial; D021122:Protein Subunits; D011993:Recombinant Fusion Proteins; D001710:Biotin