Mechanisms of assembly and cellular interactions for the bacterial genotoxin CDT.
Many bacterial pathogens that cause different illnesses employ the cytolethal distending toxin (CDT) to induce host cell DNA damage, leading to cell cycle arrest or apoptosis. CDT is a tripartite holotoxin that consists of a DNase I family nuclease (CdtB) bound to two ricin-like lectin domains (CdtA and CdtC). Through the use of structure-based mutagenesis, biochemical and cellular toxicity assays, we have examined several key structural elements of the CdtA and CdtC subunits for their importance to toxin assembly, cell surface binding, and activity. CdtA and CdtC possess N- and C-terminal nonglobular polypeptides that extensively interact with each other and CdtB, and we have determined the contribution of each to toxin stability and activity. We have also functionally characterized two key binding elements of the holotoxin revealed from its crystal structure. One is an aromatic cluster in CdtA, and the other is a long and deep groove that is formed at the interface of CdtA and CdtC. We demonstrate that mutations of the aromatic patch or groove residues impair toxin binding to HeLa cells and that cell surface binding is tightly correlated with intoxication of cultured cells. These results establish several structure-based hypotheses for the assembly and function of this toxin family.
Nesic
Dragana
D
Laboratory of Structural Microbiology, The Rockefeller University, New York, New York, USA.
Stebbins
C Erec
CE
D001427:Bacterial Toxins / Q000737:chemistry* / Q000378:metabolism / Q000633:toxicity*; D002453:Cell Cycle / Q000187:drug effects; D004247:DNA / Q000187:drug effects; D006367:HeLa Cells / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D009154:Mutation; D020836:Protein Structure, Quaternary; D013329:Structure-Activity Relationship
D001427:Bacterial Toxins; C468508:CdtA protein, Haemophilus ducreyi; C468509:CdtC protein, Haemophilus ducreyi; C067044:cytolethal distending toxin; D004247:DNA