title: Dynamic characterization of growth and gene expression using high-throughput automated flow cytometry.
abstract: Cells adjust to changes in environmental conditions using complex regulatory programs. These cellular programs are the result of an intricate interplay between gene expression, cellular growth and protein degradation. Technologies that enable simultaneous and time-resolved measurements of these variables are necessary to dissect cellular homeostatic strategies. Here we report the development of an automated flow cytometry robotic setup that enables real-time measurement of precise and simultaneous relative growth and protein synthesis rates of multiplexed microbial populations across many conditions. These measurements generate quantitative profiles of dynamically evolving protein synthesis and degradation rates. We demonstrate this setup in the context of gene regulation of the unfolded protein response (UPR) of Saccharomyces cerevisiae and uncover a dynamic and complex landscape of gene expression, growth dynamics and proteolysis following perturbations.
authors.affiliation: 1] Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, California, USA. [2] The California Institute for Quantitative Biosciences, San Francisco, California, USA.
authors.forename: Ignacio A
authors.initials: IA
authors.lastname: Zuleta
authors.affiliation: 1] Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, California, USA. [2] The California Institute for Quantitative Biosciences, San Francisco, California, USA.
authors.forename: Andrés
authors.initials: A
authors.lastname: Aranda-Díaz
authors.affiliation: 1] Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, California, USA. [2] The California Institute for Quantitative Biosciences, San Francisco, California, USA.
authors.forename: Hao
authors.initials: H
authors.lastname: Li
authors.affiliation: 1] Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, California, USA. [2] The California Institute for Quantitative Biosciences, San Francisco, California, USA.
authors.forename: Hana
authors.initials: H
authors.lastname: El-Samad
mesh_terms: D001331:Automation; D005434:Flow Cytometry / Q000295:instrumentation* / Q000379:methods*; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal / Q000502:physiology*; D012441:Saccharomyces cerevisiae / Q000254:growth & development* / Q000378:metabolism*
chemical_list: D005656:Fungal Proteins