Identification and Characterization of trans-Translation Inhibitors
Open Access
- Author:
- Davison, Alexis
- Area of Honors:
- Biochemistry and Molecular Biology
- Degree:
- Bachelor of Science
- Document Type:
- Thesis
- Thesis Supervisors:
- Ken Keiler, Thesis Supervisor
Ying Gu, Thesis Honors Advisor - Keywords:
- trans-translation
antibiotic resistance
ribosome profiling
KKL-4669
high-throughput screen
luciferase reporter assay
bacterial pathogens
Caulobacter
in vitro transcription/translation assay - Abstract:
- Individuals around the world are acquiring deadly infections from previously “common” and “non-lethal” sicknesses because common antibiotics are proving to be ineffective. The urgency for development of novel antibiotics is tremendous. In the early 20th century, antibiotics were viewed as the saving remedy to many ailments that would otherwise lead to death such as diarrhea and pneumonia. Now, the world is witnessing an increasing emergence of resistant bacterial pathogens that will cause more global deaths than any other condition by 2050.1 This work aimed at utilizing trans-translation as a target pathway for novel antibiotic development. This is an essential pathway in many bacterial pathogens that rescues ribosomes that stall during translation due to the formation of nonstop complexes. These complexes form when no stop codon is present at the 3’ end of an mRNA message being translated.3 This disables the termination of the message which is needed to produce a functional protein. If trans-translation is activated, its main machinery consisting of tmRNA and SmpB protein identifies the stalled ribosome and, through several steps, tags the protein for degradation.3 The first project featured in this work highlighted the process of identifying inhibitors of this pathway in resistant bacterial pathogens. Characterization of such inhibitors was explored to determine potential candidates for novel antibiotic development. The second project aimed to reveal molecular mechanisms occurring during trans-translation, as knowledge of this entire process has not been exhausted. This was initiated by completing a cloning project to begin ribosome profiling in a C. crescentus model. The profiling will provide information about mRNA sequences present in ribosomes while nonstop complexes are present during trans-translation and what conditions these sequences are dependent on. Therefore, continuation of both projects will aid in the movement to combat antibiotic resistance through research efforts.