Orthogonal interactome profiling of RNF41

ABSTRACT: Ring finger protein 41 (RNF41) is an E3 ubiquitin ligase involved in the ubiquitination and degradation of many proteins such as ErbB3 receptors, BRUCE and parkin. Next to this, RNF41 regulates the intracellular trafficking of certain JAK-2 associated cytokine receptors by ubiquitinating and suppressing USP8 which in turn destabilizes the ESCRT-0 complex. To further elucidate the function of RNF41 we used different orthogonal approaches to reveal the RNF41 protein complex: Affinity Purification-Mass Spectrometry, BioID and Virotrap. We combined these results with known datasets for RNF41obtained with microarray MAPPIT and Y2H screens. By combining all these datasets , we provide a comprehensive high resolution interactome network. To further remove potential methodological artifacts from this network, we further distilled the data into a high confidence interactome map by retaining protein hits identified in two or more of the orthogonal methods. AP2S1, a novel RNF41 interaction partner, was selected from this high confidence interactome for further functional validation. We reveal a role for AP2S1 in leptin and LIF receptor signaling and show that RNF41 stabilizes and relocates AP2S1.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Epithelial Cell, Cell Culture

DISEASE(S): Disease Free

SUBMITTER: Sven Eyckerman  

LAB HEAD: Sven Eyckerman

PROVIDER: PXD007794 | Pride | 2018-03-26

REPOSITORIES: Pride