Determination of hypersensitivity to genotoxic agents among Escherichia coli single gene knockout mutants.
We have tested the KEIO collection of 3985 different viable single gene knockouts in Escherichia coli to identify genes whose loss increases sensitivity to one or more of six different chemotherapeutic agents and mutagens: Bleomycin (BLM), Cisplatin (CPT), ICR-191 (ICR), 5-azacytidine (5AZ), Zebularine (ZEB), and 5-bromo-2'-deoxyuridine (5BdU). We discovered a set of 156 strains that display a significant increase in sensitivity to at least one of the agents tested. Each genotoxic agent generates a distinct "sensitivity profile" that is characteristic of the agent. Comparison with an independent study of sensitivity profiles for an extensive set of antibiotics pinpoints those effects that are relatively specific for each agent. In some cases engineered double mutants have greatly increased effects. These results provide insight into the mechanism of action of each agent, and define targets for the design of co-drugs that can potentiate these agents. An example is the finding that mutants lacking one of several genes in the folate biosynthetic pathway are hypersensitive to ZEB, leading to a demonstration of synergy between trimethoprim and ZEB.
Becket
Elinne
E
Department of Microbiology, Immunology, and Molecular Genetics, University of California-Los Angeles, CA 90095, USA.
Chen
Frank
F
Tamae
Cindy
C
Miller
Jeffrey H
JH
D004926:Escherichia coli / Q000187:drug effects* / Q000235:genetics; D055786:Gene Knockout Techniques; D008826:Microbial Sensitivity Tests; D009153:Mutagens / Q000633:toxicity*; D009154:Mutation
D009153:Mutagens