Clusters of bioactive compounds target dynamic endomembrane networks in vivo

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 108; no. 43; pp. 17850 - 17855
• Main Authors: Drakakaki, Georgia, Robert, Stéphanie, Szatmari, Anna-Maria, Brown, Michelle Q, Nagawa, Shingo, Van Damme, Daniel, Leonard, Marilyn, Yang, Zhenbiao, Girke, Thomas, Schmid, Sandra L, Russinova, Eugenia, Friml, Jiří, Raikhel, Natasha V, Hicks, Glenn R
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences 25.10.2011; National Acad Sciences
• Subjects: Algorithms; Arabidopsis - metabolism; Arabidopsis thaliana; Auxins; bioactive properties; Biological Sciences; Biological Transport - physiology; Cell Membrane - metabolism; Cells, Cultured; Cluster Analysis; data collection; drugs; Eukaryotes; eukaryotic cells; Fluorescent Antibody Technique; Freight; Genotype & phenotype; Green Fluorescent Proteins; HeLa Cells; Humans; Membranes; Microscopy, Confocal; Molecular Structure; Molecules; Nicotiana; Perceptual localization; Phenotypes; physiological transport; Plant cells; Plants; plasma membrane; Recycling; Seedlings; Seedlings - metabolism; Small Molecule Libraries - classification; Small Molecule Libraries - metabolism; Time-Lapse Imaging; Transport Vesicles - metabolism; Vacuoles
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.1108581108

Endomembrane trafficking relies on the coordination of a highly complex, dynamic network of intracellular vesicles. Understanding the network will require a dissection of cargo and vesicle dynamics at the cellular level in vivo. This is also a key to establishing a link between vesicular networks and their functional roles in development. We used a high-content intracellular screen to discover small molecules targeting endomembrane trafficking in vivo in a complex eukaryote, Arabidopsis thaliana. Tens of thousands of molecules were prescreened and a selected subset was interrogated against a panel of plasma membrane (PM) and other endomembrane compartment markers to identify molecules that altered vesicle trafficking. The extensive image dataset was transformed by a flexible algorithm into a marker-by-phenotype-by-treatment time matrix and revealed groups of molecules that induced similar subcellular fingerprints (clusters). This matrix provides a platform for a systems view of trafficking. Molecules from distinct clusters presented avenues and enabled an entry point to dissect recycling at the PM, vacuolar sorting, and cell-plate maturation. Bioactivity in human cells indicated the value of the approach to identifying small molecules that are active in diverse organisms for biology and drug discovery.