Neddylation requires glycyl-tRNA synthetase to protect activated E2

Neddylation is a post-translational modification that controls cell cycle and proliferation by conjugating the ubiquitin-like protein NEDD8 to specific targets. Here we report that glycyl-tRNA synthetase (GlyRS), an essential enzyme for protein synthesis, also plays a critical role in neddylation. In human cells, knockdown of GlyRS, but not a different tRNA synthetase, decreases the global level of neddylation and causes cell cycle abnormality. This function of GlyRS is achieved through direct interactions with multiple components of the neddylation pathway, including NEDD8, E1, and E2 (Ubc12). Using various structural and functional approaches, we show that GlyRS binds to the APPBP1 subunit of E1 to capture and protect the activated E2 (NEDD8-conjugated Ubc12) before it reaches a downstream target. Therefore, GlyRS functions as a chaperone to critically support neddylation. This function is likely to be conserved in all eukaryotic GlyRS, and may contribute to the strong association of GlyRS with cancer progression.