Mechanistic studies of inhibitors of DNA replication restart pathways in Neisseria gonorrhoeae

Date of Award

2013

Degree Name

M.S. in Chemistry

Department

Department of Chemistry

Advisor/Chair

Advisor: Matthew E. Lopper

Abstract

Complete and faithful replication of a cell's genetic information is an essential process. Many enzymes are involved in the process of successfully duplicating a cell's genetic information. Helicases, DNA polymerases, ligases, nucleases, and DNA binding proteins all play a role in DNA replication. However, the integrity of these enzymes can be compromised when they encounter DNA damage, which in general could be caused by chemical mutagens, ionizing radiations, or reactive oxidative species. Bacterial cells use a pathway called DNA replication restart" to resume DNA replication following a disruptive encounter of the DNA replication enzymes with DNA damage. This pathway is catalyzed by primosome proteins, including PriA, PriB, PriC, DnaT, DnaB, DnaC, and DnaG. The importance of DNA replication restart for bacterial cell survival is demonstrated by the inability of strains that carry mutations in key primosome genes to grow and resist DNA damaging agents. Furthermore, this pathway is specific for bacterial cells: human cells don't use the same replication restart pathway and they don't encode genes for the primosome proteins that function in bacteria. Since DNA replication restart pathways are essential for bacterial cell growth and survival and are notably absent in human cells, we seek to answer the following question: can bacterial DNA replication restart pathways be targeted with novel antibacterial compounds? In order to answer this question, we have developed an enzyme based assay for high-throughput inhibitor screening to identify compounds that block the function of the primosome proteins PriA and PriB. Several interesting lead compounds have already been identified from the preliminary screening. In this study, the lead compounds have been validated as legitimate inhibitors and characterized with respect to their potency and mechanism of action."

Keywords

DNA replication Inhibitors, Antibacterial agents Testing, Neisseria gonorrhoeae Treatment Research, Biochemistry; chemistry; replication restart pathways; Neisseria gonorrhoeae; PriA; PriB; inhibitor; inhibition

Rights Statement

Copyright © 2013, author

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