About the Project
Bacterial multidrug resistance is a growing burden on human health worldwide. Large, low copy number plasmids carrying resistance genes shuttle easily from cell to cell in bacterial populations, causing intra-species and inter-species spreading of antibiotic resistance. Multidrug resistance plasmids harbour their own survival system, a partition cassette, which ensures an accurate segregation of the plasmids from one generation to the next at cell division. When this system malfunctions, the plasmid is lost from the cell and eventually disappers from the population. The multidrug resistance plasmid TP228 replicates at low copy number in Escherichia coli. The partition cassette of TP228 consists of the parFG genes and upstream noncoding sequence parH, which harbours a series of repeats and acts as a plasmid centromere. ParF is an ATPase that assembles into a matrix through the volume of the bacterial chromosome (1). The partner protein ParG is a site-specific DNA-binding protein that associates to the plasmid at the centromere and, via interaction with ParF, recruits the plasmid to the ParF matrix. When the interplay between ParF and ParG is disrupted, the plasmid is excluded from the ParF Venus flytrap and eventually lost. This project will investigate the mechanism whereby ParG stimulates ParF ATPase activity, which leads to the restructuring of the ParF matrix.
The work will involve molecular biology and biochemical approaches in parallel with fluorescence microscopy to visualize proteins and plasmid localization in bacterial cells.
1. McLeod B, Allison-Gamble GE, Barge MT, Tonthat NK, Schumacher MA, Hayes F, Barillà D (2017) A three-dimensional ParF meshwork assembles through the nucleoid to mediate plasmid segregation. Nucleic Acids Res 45, 3158-3171
This is a self-funded project. Applicants need to have adequate funds to meet the costs of a self-funded research project including tuition fees and living expenses for the duration of the research programme. Please see information on tuition fee costs, living expenses and funding opportunities.