Observing cellular responses to perturbations is central to generating and testing hypothesis in biology. Time-lapse microscopy is a powerful tool for studying cellular phenotypes, but has been refractory to proteome-wide investigations. We developed a live-cell microarray integrated with a massively parallel microchemostat array to quantify microbial phenotypes in large-scale. Specifically, we quantified the spatio-temporal flux of the yeast proteome in response to environmental stresses by determining single-cell protein abundance and localisation changes in 4,085 GFP-tagged strains. Analysis of over 23,000 movies and 1.5 × 10^8 cells provided insight into the precise temporal orchestration of protein abundance and localisation changes in yeast. Highly dynamic re-location events are complementary to slow and persistent protein abundance changes, and together coordinate the cellular response on different time-scales. In particular, we observed that p-bodies rapidly form in response to UV-induced DNA/RNA damage. Through the precise spatio-temporal analysis of over 500 deletion - GFP-tagged strains we could determine that the p-body response is an intricate component of the DNA damage response pathway, and link it to other previously identified components. In summary, we developed a methodology for large-scale single-cell analysis and applied it to the characterisation of the stress response pathways in S. cerevisiae. Our approach is broadly applicable, and we have shown that other microbes such as E. coli, M. smegmatis, and S. pombe, can also be analysed on our platform.
Lunch @ 11:45, talk @ 12-1:00