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  • Final Activity Report Summary - SIMODSS (Single molecule dynamics of stress signalling components studied with the ulta-sensitive fluorescence microscope Cytoscout for drug screening)
FP6

SIMODSS Sintesi della relazione

Project ID: 3091
Finanziato nell'ambito di: FP6-MOBILITY
Paese: Austria

Final Activity Report Summary - SIMODSS (Single molecule dynamics of stress signalling components studied with the ulta-sensitive fluorescence microscope Cytoscout for drug screening)

The overall goal of the project was to elucidate how membrane dynamics influence the expression of genes, which are linked to pathological conditions. One prominent example in this respect are heat shock proteins (Hsps). So far, conventional biochemical techniques were used to study the machanisms of Hsp translocation to the membrane and its physical state in the membrane. However, these measurements are not capable to study membrane dynamics in detail in living cells. By contrast, single molecule techniques like single dye tracing (SDT) enable direct observation of the movement of individual molecules in cellular membranes.

The project brought together an academic research institution, providing key expertise in Hsp function (Biological Research Center of the Hungarian Academy of Sciences (BRC)) and an applied research partner focusing on exploring the potential of single molecule techniques (Upper Austrian Research GmbH (UAR)). As a first objective an ultra-sensitive fluorescence microscope in order to visualize single fluorescently labelled molecules was installed at BRC. This large area ultra-sensitive microscopy was then applied to explore timecourses of Hsp upregulation induced by physiological heat stress for a large number of cells at the level of individual cell and will in the future be applied to study variations in Hsp expression levels upon dependent on compounds added. This laid the foundation for membrane-based drug screening assays, which are based on the expertise of both host organisations. As a parallel objective a model system that allowed studying the upregulation of Hsps in cancer cells upon stress was established at UAR.

This model system was ideally suited for the available ultra-sensitive microscopy platform and allowed for the first time to decouple the effects of stress and elevated intracellular Hsp70 on the surface exposure of this chaperone. The found data are relevant for the protective and immune stimulatory role of Hsp70 exposed on the cellular surface.

Reported by

UPPER AUSTRIAN RESEARCH GMBH
Scharitzerstrasse 6-8
4020 LINZ
Austria
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