target identification and analysis of trans-translation inhibitor KKL-35

Open Access
- Author:
- Hosangadi, Divya
- Area of Honors:
- Microbiology
- Degree:
- Bachelor of Science
- Document Type:
- Thesis
- Thesis Supervisors:
- Kenneth Charles Keiler, Thesis Supervisor
Dr. Sarah Ellen Ades, Thesis Honors Advisor
Scott Brian Selleck, Faculty Reader - Keywords:
- antibiotic
trans-translation
biochemistry
E. coli - Abstract:
- Antibiotic resistance is one of the most significant public health concerns today. Compounds that target trans-translation could serve as effective broad spectrum antibiotics. trans-Translation is an error prevention mechanism for bacterial survival and virulence that involves molecules tmRNA and SmpB recognizing ribosomes translating flawed mRNA and degrading the polypeptide being formed. KKL-35 is an antibiotic discovered through high-throughput screening that targets trans-translation. This compound can strongly inhibit growth of pathogenic bacteria such as E. coli, S. flexneri, and B. anthracis at very low concentrations. I have done experimentation indicating that isolating target-based resistant mutants to KKL-35 is very difficult. Therefore, this compound is possibly less susceptible to the development of drug resistance in clinical settings. Although KKL-35 is a powerful antibiotic, how it inhibits trans-translation is unknown. In order to identify the target(s) of this compound, multiple biochemical assays were done. Techniques such as thin layer chromatography and gel electrophoresis were used to detect the presence of KKL-35 or derivatives of the compound. This research into understanding how KKL-35 works is critical for our laboratory’s goal to introduce and optimize the next generation of antibacterial therapies.