An antibody free approach to probe the presence of poly-ubiquitin chains on C. elegans sperm derived organelles after fertilization

MicroPubl Biol. 2023 Sep 19:2023:10.17912/micropub.biology.000972. doi: 10.17912/micropub.biology.000972. eCollection 2023.

Abstract

Upon C. elegans 's oocyte fertilization, the sperm brings mitochondria and membranous organelles (MOs) which are rapidly eliminated by autophagy. Their poly-ubiquitylation is suspected to be a signal for their recognition and degradation but mitochondria poly-ubiquitylation remains debated. Using fluorescent Tandem-repeated Ubiquitin-Binding Entities (TUBEs) we confirmed the presence of K48- and K63-ubiquitin chains on MOs contrasting with the absence of signal on sperm mitochondria. This new and sensitive approach confirmed the poly-ubiquitylation of the MOs while providing additional arguments for the absence of substantial poly-ubiquitylation of sperm-derived mitochondria, suggesting that K63- and K48-poly-ubiquitylation are unlikely acting as a common targeting signal for their degradation.