Mariska Twaalfhoven: Phase Separation of the Auxin Response Factors

The Auxin Response Factors (ARFs) are the transcription factors responsible for orchestrating the growth and development of land plants. How the ARFs change the expression of genes and alter the flux across molecular pathways in response to auxin remains elusive. ARFs show exciting behaviors in plants and demonstrate liquid-liquid phase separation (LLPS), a widely emerging phenomenon that equips proteins with unique abilities. What these abilities are, and what role phase separation plays in plants, is the aim of this research. The multidisciplinary approach to examine LLPS of ARFs will feature techniques from biochemistry, food physics, and fluorescence microscopy. What clues in the primary protein sequence drive phase separation will be investigated by bioinformatics. The leads will be examined by protein engineering and visualization of protein dynamics in vitro and in vivo through fluorescence microscopy and phase contrast microscopy. Membrane osmometry and NMR will be used to quantify LLPS of isolated ARFs to develop phase diagrams. The research aims to further our understanding of how protein behaviors as LLPS are fundamental for the organization and function of intracellular processes.