Completed Genetics & Molecular Biology Infection & Immunity

Investigating the physiological DNA replication initiation reaction

Funder Wellcome Charity-funded
Recipient organisation Newcastle University
Funding £1.5M
Period Oct 2017 — Oct 2022

In plain English

AI plain-English summary

Every time a bacterial cell divides, it must first copy its entire genome—and the opening move of that process, where specific proteins pry apart the DNA double helix at precise starting points, remains surprisingly mysterious. This matters because DNA replication is essential for all life, yet scientists still do not fully understand what makes a bacterial replication origin work or exactly how the initiator proteins do their job. The researcher has built a custom tool—an inducible system that swaps in foreign replication machinery—to systematically test which parts of the origin and the proteins are truly necessary inside living cells. This approach has already uncovered a previously unknown essential element in the bacterial replication origin. The work is fundamental science. It asks how a core biological machine operates, with no immediate practical application. However, bacterial DNA replication machinery is a largely untapped target for new antibiotics. A precise molecular map of the initiation reaction could eventually guide efforts to disrupt replication in pathogenic bacteria. Past fundamental discoveries about DNA replication enzymes, for example, directly enabled PCR—a technique now used daily in diagnostics, forensics, and research worldwide.

View original technical description
In all cells chromosome replication requires key initiator proteins to unwind the DNA at specific sites termed origins. Despite the fundamental importance of DNA replication initiation, crucial aspects of the process remain poorly understood. My vision is to identify all essential features of both the bacterial replication origin and DNA replication initiation proteins in vivo. This comprehensive reverse genetic analysis will guide biochemical investigations into the activities associated with deleterious mutations, thereby revealing the interactions and steps necessary for the physiological initiation reaction. Towards this goal I have created a bespoke tool: an inducible heterologous replication initiation system that allows construction and characterization of mutations within endogenous replication initiation factors. This methodology, combined with assays we developed to analyze DNA replication initiation in vitro, led us to identify a new essential bacterial replication origin element (Richardson et al. Nature 2016). We will build upon this successful approach and go on to develop more sophisticated heterologous replication systems, opening the door to studying all aspects of chromosome replication. Importantly, the bacterial DNA replication machinery is an underexploited drug target. Knowledge of bacterial DNA replication resulting from this work will provide a guide for disrupting this process in pathogenic species.

View the original record at the funder ↗

Researchers

Heath Murray (EPMC Awardee)

Related Research

Grants with similar aims, by meaning.

Assembly of the bacterial DNA replication initiation complex
Illuminating the molecular pathways of DNA replication initiation.
Bilateral NSF/BIO-BBSRC- Remodelling Replication Roadblocks: Regulatory Systems that Integrate DNA Replication, Recombination and Protein Modification
Investigating the DnaA-trio, a new essential bacterial replication origin element that specifies single-stranded DNA initiator binding
What regulates replication origin activation?

Original classification

Senior Research Fellowship Basic

Plain English summaries and category classifications on this site are generated by AI and may not perfectly reflect the original research.