Summary: Ubiquitination controls numerous cellular processes, and its deregulation is associated with many pathologies.The Nse1 subunit in the Smc5/6 complex contains a RING domain with ubiquitin E3 ligase activity and essential functions in genome integrity.However, Nse1-dependent ubiquitin targets remain elusive.Here, we use label-free quantitative proteomics to analyze the here nuclear ubiquitinome of nse1-C274A RING mutant cells.
Our results show that Nse1 impacts the ubiquitination of several proteins involved in ribosome biogenesis and metabolism that, importantly, extend beyond canonical functions of Smc5/6.In addition, our analysis suggests a connection between Nse1 and RNA polymerase I (RNA Pol I) ubiquitination.Specifically, Nse1 and the Smc5/6 complex promote ubiquitination of K408 and K410 in the clamp domain of Rpa190, a modification that induces its degradation in response to blocks in transcriptional elongation.We propose that this mechanism contributes to 5326058hx Smc5/6-dependent segregation of the rDNA array, the locus transcribed by RNA Pol I.