Dynamic characterization of growth and gene expression using high-throughput automated flow cytometry

Area of Science:

• Cell Biology
• Systems Biology
• Biotechnology

Background:

• Cells employ intricate regulatory programs involving gene expression, growth, and protein degradation to adapt to environmental changes.
• Dissecting cellular homeostatic strategies requires technologies for simultaneous, time-resolved measurements of these key variables.

Purpose of the Study:

• To develop an automated robotic system for real-time, multiplexed measurement of microbial population growth and protein synthesis rates.
• To generate quantitative profiles of dynamic protein synthesis and degradation rates under various conditions.

Main Methods:

• Development of an automated flow cytometry robotic setup.
• Real-time, simultaneous measurement of relative growth and protein synthesis rates.
• Application to study the unfolded protein response (UPR) in Saccharomyces cerevisiae.

Main Results:

• The system enables precise, time-resolved measurement of cellular growth and protein synthesis dynamics.
• Quantitative profiles of evolving protein synthesis and degradation rates were generated.
• Complex gene expression, growth, and proteolysis landscapes were uncovered in response to UPR perturbations.

Conclusions:

• The developed automated system is a powerful tool for dissecting cellular homeostatic strategies.
• This technology provides novel insights into the dynamic interplay of gene expression, growth, and protein degradation.
• The study reveals the complex regulatory landscape of the unfolded protein response in yeast.