Systematic Characterization of Fluorescent Protein Maturation in Budding Yeast.
Paolo Guerra, Luc-Alban Vuillemenot, Brady Rae, Valeriia Ladyhina, Andreas Milias-Argeitis
Author Information
Paolo Guerra: Molecular Systems Biology, Groningen Biomolecular Sciences & Biotechnology Institute, University of Groningen, 9747 AG Groningen, Netherlands.
Luc-Alban Vuillemenot: Molecular Systems Biology, Groningen Biomolecular Sciences & Biotechnology Institute, University of Groningen, 9747 AG Groningen, Netherlands.
Brady Rae: Molecular Systems Biology, Groningen Biomolecular Sciences & Biotechnology Institute, University of Groningen, 9747 AG Groningen, Netherlands.
Valeriia Ladyhina: Molecular Systems Biology, Groningen Biomolecular Sciences & Biotechnology Institute, University of Groningen, 9747 AG Groningen, Netherlands.
Andreas Milias-Argeitis: Molecular Systems Biology, Groningen Biomolecular Sciences & Biotechnology Institute, University of Groningen, 9747 AG Groningen, Netherlands. ORCID
Fluorescent protein (FP) maturation can limit the accuracy with which dynamic intracellular processes are captured and reduce the brightness of a given FP in fast-dividing cells. The knowledge of maturation timescales can therefore help users determine the appropriate FP for each application. However, maturation rates can greatly deviate from estimates that are mostly available. In this work, we present the first systematic study of maturation for 12 FPs in budding yeast. To overcome the technical limitations of translation inhibitors commonly used to study FP maturation, we implemented a new approach based on the optogenetic stimulations of FP expression in cells grown under constant nutrient conditions. Combining the rapid and orthogonal induction of FP transcription with a mathematical model of expression and maturation allowed us to accurately estimate maturation rates from microscopy data in a minimally invasive manner. Besides providing a useful resource for the budding yeast community, we present a new joint experimental and computational approach for characterizing FP maturation, which is applicable to a wide range of organisms.
