Ubiquitin selective ribosome profiling reveals systematic principles of co-translational quality control
Protein biogenesis is a stress- and error-sensitive process that can lead to nascent protein misfolding and aggregation, challenging cellular proteostasis.1 Co-translational ubiquitination (CTU) is a critical surveillance mechanism,2,3 yet its regulatory principles remain unclear due to limited known substrates and the lack of translatome-wide methods to query CTU. Here, we introduce UbSeRP, an approach that enables ubiquitin linkage-specific, and translatome-wide mapping of CTU. Focusing on ribosome-associated quality control (RQC), we expand the known endogenous RQC substrates in S. cerevisiae from few4-7 to thousands, reveal that only a subset of all identified disomes undergoes RQC, and uncover biophysical features of nascent chains that predict RQC engagement. We also identify widespread RQC-independent ubiquitination of nascent proteins, implicating broader roles of CTU in protein complex assembly. In a chronological aging model, we show that aging remodels translation and diminishes RQC engagement, favoring RQC-independent proteasomal pathways. Our findings provide systematic insight into the determinants and adaptability of CTU in maintaining proteostasis under changing physiological conditions.