Functional interaction of ubiquitin ligase RNF167 with UBE2D1 and UBE2N promotes ubiquitination of AMPA receptor

Protein ubiquitination has been historically associated with protein degradation, but recent studies have demonstrated other cellular functions substrate ubiquitination. Among the RING-type ubiquitin E3 ligase enzymes present in human genome, RNF167 is a transmembrane located endosomes and lysosomes implicated controlling endolysosomal pathway. Substrates of identified, ubiquitin-conjugating E2 involved mechanism remain unknown. In this study, we describe interaction between conjugating enzymes. By means vitro autoubiquitination binding assays, show that functionally interacts many E2s, while fluorescence microscopy illustrates these interactions occur lysosomes. Kinetic analyses selected reveal submicromolar dissociation constants. The computed model RING domain gives us insights on how could interact Furthermore, results polyubiquitination AMPA-type glutamate receptor subunit GluA2, one RNF167’s known substrates, possible by enzyme UBE2N only after GluA2 primed subsequent to action an initiating RNF167. Pharmacological inhibition cultured hippocampal neurons diminishes AMPA-induced This study characterizes interacting partners constitutes initial step toward identification functional pairs assembled from substrate.