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Infection and Immunity, September 2003, p. 4883-4890, Vol. 71, No. 9
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.9.4883-4890.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Interactions of Campylobacter jejuni Cytolethal Distending Toxin Subunits CdtA and CdtC with HeLa Cells

Robert B. Lee, Duane C. Hassane, Daniel L. Cottle, and Carol L. Pickett*

Department of Microbiology, Immunology, and Molecular Genetics, School of Medicine, University of Kentucky, Lexington, Kentucky 40536

Received 21 February 2003/ Returned for modification 11 April 2003/ Accepted 16 June 2003

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 G1 or G2 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.

* Corresponding author. Mailing address: Department of Microbiology, Immunology, and Molecular Genetics, University of Kentucky Medical Center, 800 Rose St., MS 415, Lexington, KY 40536-0298. Phone: (859) 323-5313. Fax: (859) 257-8994. E-mail: cpicket{at}uky.edu.

Editor: J. T. Barbieri

Infection and Immunity, September 2003, p. 4883-4890, Vol. 71, No. 9
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.9.4883-4890.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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