RNF41 interacts with the VPS52 subunit of the GARP and EARP complexes

Abstract

RNF41 (Ring Finger Protein 41) is an E3 ubiquitin ligase involved in the intracellular sorting and function of a diverse set of substrates. Next to BRUCE and Parkin, RNF41 can directly ubiquitinate ErbB3, IL-3, EPO and RARα receptors or downstream signaling molecules such as Myd88, TBK1 and USP8. In this way it can regulate receptor signaling and routing. To further elucidate the molecular mechanism behind the role of RNF41 in intracellular transport we performed an Array MAPPIT (Mammalian Protein-Protein Interaction Trap) screen using an extensive set of proteins derived from the human ORFeome collection. This paper describes the identification of VPS52, a subunit of the GARP (Golgi-Associated Retrograde Protein) and the EARP (Endosome-Associated Recycling Protein) complexes, as a novel interaction partner of RNF41. Through interaction via their coiled coil domains, RNF41 ubiquitinates and relocates VPS52 away from VPS53, a common subunit of the GARP and EARP complexes, towards RNF41 bodies.

CACYBP Enhances Cytoplasmic Retention of P27Kip1 to Promote Hepatocellular Carcinoma Progression in the Absence of RNF41 Mediated Degradation

“…RNF41 recruits E2 conjugating enzymes with its N-terminal RING finger domain (residues 18-57), followed by a coiled-coil domain essential for homotrimerization (residues 135-179), and interacts with the substrates via its C-terminal domain (residues 193-317) 17, 19. In addition to promoting degradation, ubiquitination by RNF41 also controls the sorting, processing and subcellular relocalization of the substrates, as demonstrated for LR (leptin receptor), LIFR (leukemia inhibitory factor receptor), IL-6R (interleukin-6 receptor) 20, USP8 21 and VPS52 22. RNF41 exerts differential ubiquitination functions towards Myd88 (poly-lys48) and TBK1 (poly-lys63) and regulates TLR-mediated production of type-I interferon 23.…”

VPS51 biallelic variants cause microcephaly with brain malformations: A confirmatory report

“…GARP differs only in one subunit with another tethering complex, known as the endosome-associated recycling protein (EARP) complex, in which the VPS54 subunit is substituted by syndetin (VPS50). EARP ensures the fusion of endosome-derived carriers with recycling endosomes and therefore promotes recycling of endocytic membranes back to the plasma membrane (Masschaele et al, 2017;Schindler et al, 2015).…”

Straightforward Protein-Protein Interaction Interface Mapping via Random Mutagenesis and Mammalian Protein Protein Interaction Trap (MAPPIT)

“…We successfully applied random mutagenesis combined with MAPPIT for the interaction interface analysis of six targets in four unrelated protein families: the antiviral host restriction factor apolipoprotein B messenger RNA-editing catalytic polypeptide-like G (Apobec3G) [22], the ring finger protein 41 (RNF41) [23], the ligand binding domain of peroxisome proliferator-activated receptor α (PPAR-α) and the Toll/IL-1R (TIR) domains of the Toll-like receptor adapters MyD88 adapter-like (Mal) [18], myeloid differentiation primary response gene 88 (MyD88) [19] and TIR-domain-containing adapter inducing interferon-β (TRIF). For five of these targets, we tested the interaction of the mutant libraries with three or four different interactors in parallel.…”