The Arabidopsis ESCRT protein–protein interaction network

• DOI: 10.1007/s11103-011-9770-4
• Publication Date: 2011-03-25T22:21:00Z
• Authors: Mojgan Shahriari; Klaus Richter; Channa Keshavaiah; Aneta Sabovljevic; Martin Huelskamp; Swen Schellmann
• Keywords: ESCRT; Bimolecular fluorescence complementation; Transport protein; Two-hybrid screening; Interactome

In yeast, endosomal sorting of monoubiquitylated transmembrane proteins is performed by a subset of the 19 "class E vacuolar protein sorting" proteins. The core machinery consists of 11 proteins that are organised in three complexes termed ESCRT I-III (endosomal sorting complex required for transport I-III) and is conserved in eukaryotic cells. While the pathway is well understood in yeast and animals, the plant ESCRT system is largely unexplored. At least one sequence homolog for each ESCRT component can be found in the Arabidopsis genome. Generally, sequence conservation between yeast/animals and the Arabidopsis proteins is low. To understand details about participating proteins and complex organization we have performed a systematic pairwise yeast two hybrid analysis of all Arabidopsis proteins showing homology to the ESCRT core machinery. Positive interactions were validated using bimolecular fluorescence complementation. In our experiments, most putative ESCRT components exhibited interactions with other ESCRT components that could be shown to occur on endosomes suggesting that despite their low homology to their yeast and animal counterparts they represent functional components of the plant ESCRT pathway.