CELL CYCLE PALM · Quantifying regulatory protein diffusion in the bacterial cell cycle by high-throughput single particle tracking PALM
7РП — „Хора“ (Действия „Мария Кюри“)
- Период
- 2012-09-01 → 2014-08-31
- Финансиране от ЕС
- 192 622 €
- Участници
- 1
- Схема
- MC-IEF
Линиите свързват координатора с партньорите.
Накратко на български
Разположението на протеина FtsZ при деленето на бактериите се проследява чрез свръхразрешаваща микроскопия. По-доброто разбиране на бактериалната биология помага за откриването на нови мишени за създаване на антибиотици.
Кратко обяснение, генерирано от езиков модел по текста на CORDIS. Оригиналът е по-долу.
Резултати накратко
Quantifying regulatory protein diffusion in the bacterial cell cycle by high-throughput single particle tracking PALM
The rise of antibiotic resistance represents a grave challenge for mankind. With no new classes of antibiotics discovered since 1987, an improved understanding of bacterial cell biology will be crucial to identifying and studying novel antibiotic targets. Furthermore, bacteria represent an excellent model system for fundamental research, due to their genetic tractability and compatibility with high-throughput multiplexing. During the period of this FP7 project, we used the new technique of super-resolution fluorescence microscopy to study bacterial cell biology. This combination is extremely exciting, since it allows one of the key challenges to studying living bacteria – their small size (a few microns) – to be overcome, revealing the nanoscale organization of key bacterial proteins. Our major innovation was to conceive of and build a high-throughput super-resolution imaging modality that would allow us to study hundreds of cells in a single experiment (Fig. 1). This allowed us to study the cell-cycle dependent organization of the essential cell division protein FtsZ, a key next-generation antibiotic target. We found that FtsZ forms a patchy “band” at mid-cell (Fig. 2-3), and not a continuous ring as previously thought (Holden et al., PNAS 2014). This challenges the “FtsZ-centric” model of cell division, suggesting that FtsZ probably does not generate constrictive force, and may merely recruit other proteins to the division site. This has significant implications for our understanding of cell division, and will motivate further research into alternative physical mechanisms for constriction. The project generated multiple significant publications. In addition to a PNAS paper on FtsZ cytokinesis (Holden et al., PNAS 2014), the project also lead to or supported publications on nanoscale analysis of bacterial transcription (Endesfelder et al, Biophys. J 2013), and high speed in vivo super-resolution imaging (Min et al. Sci. Rep. 2014, Min et al. Biomed. Opt. Expr. 2014). We have also released several pieces of software related to the project, which are linked to from http://leb.epfl.ch/software. Outreach was performed by blogging (http://seamusholden.wordpress.com/), release of Youtube videos to explain the results (see website), and liasing with press (HTPALM was featured in Nature Methods and Microscopy & Microanalysis. The new results and techniques developed during the project will support further investigation and understanding of medically relevant processes in bacterial cell biology, especially bacterial cell division. This new findings will inform and support the development of new antibiotics targeting these processes.
Текст от CORDIS, на английски · Данни: CORDIS, © Европейски съюз
Цел на проекта
The precise timing and spatial organisation of the cell cycle is essential to the survival of any organism. Caulobacter crescentus is a simple, easily synchronisable organism which provides an excellent model system for cell cycle regulation, where a complex interacting network of regulatory proteins controls the precise timing and position of cell cycle processes. Although significant work has been carried out to determine the temporal and spatial dynamics of these proteins, the biophysical mechanisms by which bacterial regulatory proteins localise to specific cellular structures is largely unknown, due to the experimental challenge of studying diffusion of high-copy-number proteins.It has recently become possible to study the diffusion of high-copy-number proteins using single particle tracking photoactivated localisation microscopy (sptPALM). We will develop a high-throughput implementation of sptPALM (HT-PALM), capable of automatically recording sptPALM data for multiple cells over the duration of the cell cycle. We will use HT-PALM to study C. crescentus cell cycle regulatory proteins, determining the spatio-temporal variation in diffusion coefficient and molecular confinement (“molecular mobility”) and its effect on protein localisation dynamics.This study will advance our biophysical understanding of the role of molecular mobility in the bacterial cell cycle and at the same time provide the technological basis for a broadly useful high-throughput modality of the PALM technique. Thus, the proposed work will enable high-throughput super-resolution microscopy studies in additional prokaryotic systems, and pave the way for eukaryotic high-throughput super-resolution microscopy.""
Оригинален текст от CORDIS (на английски).
Участници
- ECOLE POLYTECHNIQUE FEDERALE DE LAUSANNE · LausanneКоординаторШвейцария
Връзки
Данни: CORDIS, © Европейски съюз