Multi-scale characterization of plant clathrin-mediated endocytosis

Session

Session 2 - Early Career Researchers Session: the Science of Tomorrow Today

Authors

Alexander Johnson (1)

Affiliations

1. Medical University of Vienna

Abstract text

Clathrin mediated endocytosis (CME) plays an important role for many physiological processes in plants, ranging from intra- and intercellular signalling, growth and development to nutrient uptake and pathogen defense. Despite this physiological significance, little is known about its molecular mechanisms. By overcoming key optical and sample preparation challenges provided by plant samples, we optimized and developed a range of quantitative imaging tools to directly examine plant CME at multiple scales; (i) in whole tissues (confocal microscopy), (ii) live single events (total internal reflection fluorescence [TIRF] and structured illumination microscopy [SIM]), and (iii) at 3D ultra-structural resolutions (scanning transmission electron microscopy [STEM] tomography). By using these tools to directly examine plant CME, and the proteins driving it, we have updated our working models of plant CME. For example, in contrast to predictions, and despite the high turgor pressure in plant cells, we found plant CME was independent of actin. We found that endocytic membrane bending was instead mediated by a plant specific protein (TPLATE – absent from mammalian and yeast genomes), highlighting that plant CME is mechanistically and evolutionary distinct from other eukaryotic model systems. I am continuing to develop novel ways of imaging plants, specifically looking to apply Brillouin light scattering approaches to define the biophysical properties of plant cells and endocytic events.