European Companies Search Engine
UK funding (£498,866): The bacterial type IV pilus machinery as a DNA translocator Ukri27 Jun 2018 UK Research and Innovation, United Kingdom
Overview
Text
The bacterial type IV pilus machinery as a DNA translocator
| Abstract | Bacteria are single-celled organisms that are surrounded by membranes made of lipids. Membranes maintain the shape and structure of the cell and also act as a semi-permeable barrier, allowing the exchange of small nutrients and waste products, but preventing leakage of larger molecules like DNA and proteins. However, many molecules need to cross membranes in order to reach the destination where they perform their function, which may be in DNA replication, protein synthesis or even as part of the "engine" of the cell, which uses energy to turn a motor and drive bacterial movement. To cross membrane barriers, many different systems exist. The overarching theme of my research is the study of a large and powerful protein complex called the type IV pilus (T4P) assembly machinery, which performs two completely different functions. The first is transporting small protein subunits across the cell membrane to produce an extremely long filament (a pilus) on the cell surface. The pilus is very dynamic and can be assembled and retracted extremely quickly, enabling cells to "walk" across surfaces in a jerky manner. The pilus also acts as a means of communication between cells, and can be a key factor in enabling bacteria to cause disease. In our previous work, we have identified two different forms of pilus that are assembled from the same machine, but it is not clear how they are related to bacterial behaviour. It is imperative that we further our knowledge of pili by conducting fundamental biosciences research. In this way, mechanisms that prevent bacterial movement and colonisation can be developed by more applied research strategies in the future. The second function of some T4P machines is their ability to support DNA uptake from the extracellular environment. It is not entirely clear if this also involves the pilus filament in some form. Uptake of DNA is called "natural competence" and can be an extremely dangerous phenomenon as it enables cells to "feed" on foreign DNA and incorporate it into their own genome. This means that previously non-risk species can behave unpredictably and become pathogenic to animals, crops and humans. By the same mechanism, bacteria can develop antimicrobial resistance, which is a massive worldwide healthcare problem. We plan to develop an assay involving fluorescent labelling, that will enable DNA binding and uptake to be monitored in whole cells, which in the subsequent part of this work will lead on to identifying the specific binding site and proteins involved. This is an extremely important aspect of research that must be conducted in order to better understand the underlying mechanisms that bacteria use to take up genetic material. The main technique that will be employed in this research is electron cryo-tomography (cryoET). This is a state-of-the-art approach, which employs an electron microscope in order to visualise whole cells and determine protein structures, which is extremely powerful in the burgeoning sphere of structural cell biology. Using cryoET in this study will allow us to visualise the entire T4P machinery performing different functions in bacterial cells. The work will therefore uncover new information to further our understanding of a fundamental molecular machine, and how it performs a dual-function in nature. |
| Category | Research Grant |
| Reference | BB/R008639/1 |
| Status | Closed |
| Funded period start | 27/06/2018 |
| Funded period end | 01/12/2022 |
| Funded value | £498,866.00 |
| Source | https://gtr.ukri.org/projects?ref=BB%2FR008639%2F1 |
Participating Organisations
| UNIVERSITY OF EXETER | |
| Alan Turing Institute | |
| ETH Zurich | |
| University of Electro-Communications | |
| Goethe University Frankfurt | |
| Newcastle University | |
| University of Bristol | |
| UNIVERSITY OF MANCHESTER |
The filing refers to a past date, and does not necessarily reflect the current state. The current state is available on the following page: University OF Exeter, Exeter.
The visualizations for "University of Exeter - UK funding (£498,866): The bacterial type IV pilus machinery as a DNA translocator"
are provided by
North Data
and may be reused under the terms of the
Creative Commons CC-BY license.
