Revealing the BActerial Response to Temperature and Antibiotics Stress via multi...
Revealing the BActerial Response to Temperature and Antibiotics Stress via multiscale imaging
Antibiotic resistance is one of the biggest scientific and health challenges of our time. If actions are not taken, antibiotics resistance infections will become the first cause of death by 2050. One of the most common signs of ba...
Antibiotic resistance is one of the biggest scientific and health challenges of our time. If actions are not taken, antibiotics resistance infections will become the first cause of death by 2050. One of the most common signs of bacterial infection is a rapid increase of the host temperature that exposes bacteria to a heat shock (HS). To survive and quickly adapt to this new environment, bacteria activate the synthesis of specific proteins, called heat shock proteins (HSPs). In E.coli, HSPs expression is mediated by the alternative sigma factor sigma32 (Sig32). The molecular processes that are associated with the bacterial stress responses induced by a combination of change in temperature and presence of antibiotics have not yet been investigated in detail due to the technical challenges of monitoring this combination of stresses with the necessary temporal and spatial accuracy. Cell-to-cell fluctuations in protein expression could affect single cells survival. Therefore, these processes should be studied with single cell precision. I propose to investigate bacterial responses to the combined stresses of temperature increase and antibiotics at single cell level in live E.coli using a combination of microfluidics and single molecule/super-resolution imaging. I will design and build a microfluid system able to control the stress landscape of temperature and antibiotic concentrations. This will allow me to study Sig32 expression during HS using fast-maturating fluorescent protein fusions. Next, I will monitor how HS affects proteins synthesis by measuring the diffusion changes of single ribosomal subunits (S30 and S50) during HS, using single particle tracking PALM. Finally, I will quantify the interplay between HS response and antibiotic susceptibility by monitoring Sig32 levels and the diffusion of ribosomal subunits. Taken together my results will provide for the first time a quantitative picture and a new framework to study bacterial response to multiple stresses.ver más
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