Clusters of bioactive compounds target dynamic endomembrane networks in vivo

Proc Natl Acad Sci U S A. 2011 Oct 25;108(43):17850-5. doi: 10.1073/pnas.1108581108. Epub 2011 Oct 17.

Abstract

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Algorithms*
  • Arabidopsis / metabolism*
  • Biological Transport / physiology
  • Cell Membrane / metabolism*
  • Cells, Cultured
  • Cluster Analysis
  • Fluorescent Antibody Technique
  • Green Fluorescent Proteins
  • HeLa Cells
  • Humans
  • Microscopy, Confocal
  • Molecular Structure
  • Seedlings / metabolism
  • Small Molecule Libraries / classification
  • Small Molecule Libraries / metabolism*
  • Time-Lapse Imaging
  • Tobacco
  • Transport Vesicles / metabolism*

Substances

  • Small Molecule Libraries
  • Green Fluorescent Proteins