MUF1/leucine-rich repeat containing 41 (LRRC41), a substrate of RhoBTB-dependent cullin 3 ubiquitin ligase complexes, is a predominantly nuclear dimeric protein

Schenková, Kristína; Lutz, Julia; Kopp, Marion; Ramos, Sonia; Rivero, Francisco

Abstract

RhoBTB (BTB stands for broad-complex, tramtrack, bric à brac) proteins are tumor suppressors involved in the formation of cullin 3 (Cul3)-dependent ubiquitin ligase complexes. However, no substrates of RhoBTB-Cul3 ubiquitin ligase complexes have been identified. We identified MUF1 (LRRC41, leucine-rich repeat containing 41) as a potential interaction partner of RhoBTB3 in a two-hybrid screening on a mouse brain cDNA library. MUF1 is a largely uncharacterized protein containing a leucine-rich repeat and, interestingly, a BC-box that serves as a linker in multicomponent, cullin 5 (Cul5)-based ubiquitin ligases. We confirmed the interaction of MUF1 with all three mammalian RhoBTB proteins using immunoprecipitation. We characterized MUF1 in terms of expres sion profile and subcellular localization, the latter also with respect to RhoBTB proteins. We found out that MUF1 is a ubiquitously expressed nuclear protein that, upon coexpression with RhoBTB, partially retains in the cytoplasm, where both proteins colocalize. We also show that MUF1 is able to dimerize similarly to other leucine-rich repeat-containing proteins. To explore the significance of MUF1-RhoBTB interaction within Cul-ligase complexes and the mechanism of MUF1 degradation, we performed a protein stability assay and found that MUF1 is degraded in the proteasome in a Cul5-independent manner by RhoBTB3-Cul3 ubiquitin ligase complex. Finally, we explored a possible heterodimerization of Cul3 and Cul5 and indeed discovered that these two cullins are capable of forming heterodimers. Thus, we have identified MUF1 as the first substrate for RhoBTB-Cul3 ubiquitin ligase complexes. Identification of substrates of these complexes will result in better understanding of the tumor suppressor function of RhoBTB.

Citation

Schenková, K., Lutz, J., Kopp, M., Ramos, S., & Rivero, F. (2012). MUF1/leucine-rich repeat containing 41 (LRRC41), a substrate of RhoBTB-dependent cullin 3 ubiquitin ligase complexes, is a predominantly nuclear dimeric protein. Journal of Molecular Biology, 422(5), 659-673. https://doi.org/10.1016/j.jmb.2012.06.016

• Journal Article Type: Article
• Acceptance Date: Jun 10, 2012
• Online Publication Date: Jun 15, 2012
• Publication Date: Oct 5, 2012
• Deposit Date: Sep 21, 2021
• Journal: Journal of Molecular Biology
• Print ISSN: 0022-2836
• Publisher: Elsevier
• Peer Reviewed: Peer Reviewed
• Volume: 422
• Issue: 5
• Pages: 659-673
• DOI: https://doi.org/10.1016/j.jmb.2012.06.016
• Keywords: Atypical Rho GTPase; Tumor suppressor; Substrates; Degradation; Cullin-dependent ubiquitin ligase
• Public URL: https://hull-repository.worktribe.com/output/540914