# A Novel High-Throughput Exploration of DNA Supercoiling toward the Therapeutic Manipulation of Bacteria

> **NIH NIH R21** · YALE UNIVERSITY · 2024 · $209,375

## Abstract

PROJECT SUMMARY
Bacterial survival requires a nearly constant and specific global DNA supercoiling status.
This is achieved by the opposing functions of the DNA-relaxing topoisomerase I and
DNA-compacting DNA gyrase. We propose the first unbiased high-throughput
exploration of genes governing the abundance and activity of these enzymes, which
hold great untapped promise for therapeutic manipulation. How pathogens change
DNA supercoiling during infection conceals vulnerabilities critical to overcoming the
toxic effects of current DNA gyrase inhibitors and emergence of bacteria resistant to
these agents. We will examine mutants of the pathogen Salmonella enterica serovar
Typhimurium for altered DNA supercoiling in infection-relevant conditions by deploying
Fluorescence Evaluation of DNA Supercoiling – or FEDS – a new high-throughput
method developed in our laboratory that reports the global DNA supercoiling status of
living bacteria. To expose the genetic basis for altered DNA supercoiling, we will identify
the mutated genes and explore genetic interactions among the mutations to reveal
novel instances of synthetic lethality, thereby uncovering both novel genetic pathways
and novel ways to manipulate DNA supercoiling. We will solve the bases for the DNA
supercoiling behavior of the identified mutants by determining the amounts of DNA
gyrase and topoisomerase I as well as the abundance of adenosine triphosphate,
adenosine diphosphate, and specific polyamines – all critical regulators of DNA gyrase
activity. The proposed research program pioneers the physiological understanding of an
essential cellular property exploitable in a wide range of bacterial species to identify
novel targets for antibacterial agents.

## Key facts

- **NIH application ID:** 10843186
- **Project number:** 5R21AI173914-02
- **Recipient organization:** YALE UNIVERSITY
- **Principal Investigator:** Eduardo Groisman
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $209,375
- **Award type:** 5
- **Project period:** 2023-05-17 → 2025-04-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10843186

## Citation

> US National Institutes of Health, RePORTER application 10843186, A Novel High-Throughput Exploration of DNA Supercoiling toward the Therapeutic Manipulation of Bacteria (5R21AI173914-02). Retrieved via AI Analytics 2026-05-31 from https://api.ai-analytics.org/grant/nih/10843186. Licensed CC0.

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