Year: 2024
Type: conference-abstract
Abstract: The Escherichia coli genome contains 5-methylcytosine at CCWGG sites.In eukaryotes, the role of DNA methylation is well-defined as an integral part of transcription regulation.However, the role of bacterial cytosine DNA methylation is less understood, with our laboratory's previous studies suggesting that cytosine DNA methylation is important for the bacterial stress response.To better understand the function of cytosine DNA methylation in bacteria, we challenged wild-type and cytosine DNA methylation mutant bacteria with antibacterial compounds to stimulate a stress response in bacteria.We also blocked cytosine DNA methylation with inhibitors to elucidate the role of cytosine DNA methylation.The novel DNA methylation inhibitor and antitumor drug, CM-272, demonstrated success in inducing DNA demethylation and disrupting cell cycle progression in cancer cells.We treated Escherichia coli cells with CM-272 to block cytosine DNA methylation and simultaneously measured bacterial growth.An impact of CM-272 on E. coli growth was not expected, as cytosine DNA methylation is not required for E. coli growth.However, CM-272 generated a zone of inhibition in wild-type Escherichia coli, E. coli cytosine DNA methylation mutants, and Bacillus subtilis, in Kirby-Bauer disk diffusion assays demonstrating that CM-272 has antimicrobial properties.Minimum inhibitory concentration assays and growth curve experiments were performed to provide quantitative results for the impact of CM-272 on bacterial growth, and also indicate that CM-272 inhibits bacterial growth.Surprisingly, CM-272 blocked bacterial growth in E. coli strains that lack a cytosine DNA methylation pathway, indicating a novel, DNA methylation-independent mechanism of action.Currently, experiments are being performed to better understand the mechanism by which CM-272 inhibits growth in both gramnegative and gram-positive bacteria and this will provide insight into both the antimicrobial properties of CM-272 and the unknown role of bacterial cytosine DNA methylation.This work was supported by the GeneseoFoundation.
Source: Journal of Biological Chemistry
Institution: University of North Dakota
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $2,800
OpenAlex ID: W4393167380