Objective Most microorganisms lack homeothermic regulation which subjects them to unpredictable fluctuations in environmental temperature. Nevertheless, many bacteria and archaea can grow across a large range of up to 40°C. Most biochemical reactions in bacteria occur in the highly crowded cytoplasm constituting a complex and dynamic interaction network of a cell. Temperature affects the rate of both intra- and intermolecular reactions, and large-scale perturbations by temperature could be disastrous to cellular function, homeostasis, and sub-cellular organization. It is still largely unknown, how temperature affects the properties of the cytoplasm and what the consequences to cellular processes are. The driving hypothesis of this project is that bacteria can actively modulate their cytoplasmic state to avoid the detrimental effects of temperature fluctuations.To test this, I have established cutting-edge super-resolution single-molecule tracking tools to directly observe molecule dynamics in live bacteria in real time. First, we will quantify the diffusion and activity of macromolecules and other probes as a function of temperature to uncover the changes in the cytoplasmic state. Second, we will probe different bacteria growing at temperatures from 0°C up to 100°C to characterize evolutionary differences in the cytoplasmic dynamics and temperature scaling of reaction rates. Finally, we will uncover mechanisms by which bacteria obtain different cytoplasmic properties. Overall, these approaches will reveal how the cytoplasmic state in bacteria changes with temperature and how this contributes to the cellular processes, which has significant implications on how microorganisms adapt to temperatures and what are the limits of cellular life. Fields of science natural sciencesbiological sciencesmicrobiologybacteriologynatural sciencesphysical sciencesopticsmicroscopysuper resolution microscopymedical and health sciencesbasic medicinephysiologyhomeostasis Keywords single-molecule tracking extremophile cytoplasm Programme(s) HORIZON.1.1 - European Research Council (ERC) Main Programme Topic(s) ERC-2022-STG - ERC STARTING GRANTS Call for proposal ERC-2022-STG See other projects for this call Funding Scheme HORIZON-ERC - HORIZON ERC Grants Host institution HELSINGIN YLIOPISTO Net EU contribution € 1 792 125,00 Address YLIOPISTONKATU 3 00014 Helsingin Yliopisto Finland See on map Region Manner-Suomi Helsinki-Uusimaa Helsinki-Uusimaa Activity type Higher or Secondary Education Establishments Links Contact the organisation Opens in new window Website Opens in new window Participation in EU R&I programmes Opens in new window HORIZON collaboration network Opens in new window Total cost € 1 792 125,00 Beneficiaries (1) Sort alphabetically Sort by Net EU contribution Expand all Collapse all HELSINGIN YLIOPISTO Finland Net EU contribution € 1 792 125,00 Address YLIOPISTONKATU 3 00014 Helsingin Yliopisto See on map Region Manner-Suomi Helsinki-Uusimaa Helsinki-Uusimaa Activity type Higher or Secondary Education Establishments Links Contact the organisation Opens in new window Website Opens in new window Participation in EU R&I programmes Opens in new window HORIZON collaboration network Opens in new window Total cost € 1 792 125,00