Proteomic analyses of the Arabidopsis cap-binding complex define core set and TOR-dependent protein components

Abstract

The eukaryotic cap-binding complex (CBC) is a hub for regulations affecting mRNA behaviour including translation, degradation and storage. Beside the core eukaryotic translation initiation factors, other proteins, many of which are yet unknown, are thought to interact stably or transiently with the CBC depending on cell status. The prototype of these regulators is the animal eIF4E binding protein (4E-BP), a direct target of the TOR (Target of Rapamycin) kinase that competes with the cap-binding protein eIF4E, thus repressing translation. In plants, no functional homologs of 4E-BP have so far been characterized. In this work we performed several deep proteomic analyses of the Arabidopsis CBC after cap-affinity purification from wild-type plants. We also investigated the CBC in eIF4E mutant plants, Arabidopsis lines with lower TOR activity, or during infection with eIF4E-dependent potyviruses, conditions which are all affecting translation at the initiation level. These analyses allowed us to define a limited core set of CBC components, which were detected in all samples. Interestingly, we identified proteins, like AGO1 or VCS, which were always detected in conditions where either TOR or mRNA translation were reduced. Meta-analysis of these data revealed several new plant interactors of the CBC, potentially defining pathways related to mRNA stability and degradation, metabolism and viral life cycle. A search for eIF4E binding motifs identified several new potential 4E-BP relatives in plants.