Bacterial resistance to antibiotics has emerged as a considerable threat to human health. Methicillin- resistant Staphloccous aureus (MRSA) and vancomycin-resistant enterococci (VRE) are multi-drug resistant bacteria that cause life threatening infections in the hospital setting and in some cases have leapt to the larger community. It is clear that new strategies and tactics are needed to combat these insidious bacteria. Often times, bacteria owe their antibiotic resistance to the proteins encoded by plasmids that they harbor. Plasmids are small, circular, extra-chromosomal pieces of DNA that can be transferred from one bacterium to another. These plasmids often contain genes that encode proteins that confer resistance to a wide array of antibiotics. Indeed, for several classes of antibiotics (including beta-lactams, macrolides, and aminoglycosides) plasmid-borne resistance is ubiquitous. In addition, many of the worst multi-drug resistant bacteria (including MRSA and VRE) are resistant by virtue of the plasmid they harbor. Proposed herein is a strategy to attack this plasmid-encoded resistance through the creation of """"""""anti-plasmid"""""""" agents, small molecules that will vanquish the plasmid from the cell, thus rendering the bacteria sensitive to antibiotics. These compounds are designed to mimic a known, naturally occurring mechanism for plasmid elimination, known as plasmid incompatibility. The bio-molecules that determine plasmid incompatibility are typically small pieces of RNA and DNA iterons. It has been shown that genetic mutation of these RNA incompatibility determinants disrupts RNA loop-loop interactions and leads to plasmid elimination.
In Specific Aims 1 and2 of this proposal, small molecules are described that will disrupt the RNA loop-loop interaction in a completely analogous manner, thus leading to plasmid elimination.
Specific Aims 3 and 4 describe studies on the mechanism and inhibition of the plasmid replication initiation protein, RepA. The successful completion of the experiments described herein could lead to a dramatic change in the manner in which antibiotic resistant infections are treated.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
3R01GM068385-05S1
Application #
7390029
Study Section
Bio-Organic and Natural Products Chemistry Study Section (BNP)
Program Officer
Fabian, Miles
Project Start
2003-04-15
Project End
2008-03-31
Budget Start
2007-05-01
Budget End
2008-03-31
Support Year
5
Fiscal Year
2007
Total Cost
$51,538
Indirect Cost
Name
University of Illinois Urbana-Champaign
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
041544081
City
Champaign
State
IL
Country
United States
Zip Code
61820
Larson, Amy S; Hergenrother, Paul J (2014) Light activation of Staphylococcus aureus toxin YoeBSa1 reveals guanosine-specific endoribonuclease activity. Biochemistry 53:188-201
van Rensburg, Julia J; Hergenrother, Paul J (2013) Detection of endogenous MazF enzymatic activity in Staphylococcus aureus. Anal Biochem 443:81-7
Williams, Julia J; Hergenrother, Paul J (2012) Artificial activation of toxin-antitoxin systems as an antibacterial strategy. Trends Microbiol 20:291-8
Halvorsen, Elizabeth M; Williams, Julia J; Bhimani, Azra J et al. (2011) Txe, an endoribonuclease of the enterococcal Axe-Txe toxin-antitoxin system, cleaves mRNA and inhibits protein synthesis. Microbiology 157:387-97
Williams, Julia J; Halvorsen, Elizabeth M; Dwyer, Ellen M et al. (2011) Toxin-antitoxin (TA) systems are prevalent and transcribed in clinical isolates of Pseudomonas aeruginosa and methicillin-resistant Staphylococcus aureus. FEMS Microbiol Lett 322:41-50
Williams, Julia J; Hergenrother, Paul J (2008) Exposing plasmids as the Achilles'heel of drug-resistant bacteria. Curr Opin Chem Biol 12:389-99
Thomas, Jason R; Hergenrother, Paul J (2008) Targeting RNA with small molecules. Chem Rev 108:1171-224
Wang, Nora R; Hergenrother, Paul J (2007) A continuous fluorometric assay for the assessment of MazF ribonuclease activity. Anal Biochem 371:173-83
Moritz, Elizabeth M; Hergenrother, Paul J (2007) Toxin-antitoxin systems are ubiquitous and plasmid-encoded in vancomycin-resistant enterococci. Proc Natl Acad Sci U S A 104:311-6
Hergenrother, Paul J (2006) Obtaining and screening compound collections: a user's guide and a call to chemists. Curr Opin Chem Biol 10:213-8

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