Reconstitution of a Minimal Ribosome-Associated Ubiquitination Pathway with Purified Factors》.pdf

Molecular Cell Article Reconstitution of a Minimal Ribosome-Associated Ubiquitination Pathway with Purified Factors Sichen Shao1 and Ramanujan S. Hegde1,* 1MRC Laboratory of Molecular Biology, Cambridge, CB2 0QH, UK *Correspondence: rhegde@mrc-lmb.cam.ac.uk /10.1016/j.molcel.2014.07.006 This is an open access article under the CC BY license (/licenses/by/3.0/). SUMMARY nascent chain ubiquitination on the ribosome has only recently been undertaken (Duttler et al., 2013; Wang et al., 2013), and Ribosomes stalled on aberrant mRNAs engage qual- the pathways are poorly understood. The best defined of these ity control mechanisms that degrade the partially is a ribosome-associated quality control (RQC) pathway that translated nascent polypeptide. Ubiquitination of degrades the partially synthesized protein products of transla- the nascent protein is mediated by the E3 ligase Lis- tionally stalled ribosomes (Bengtson and Joazeiro, 2010; Brand- terin via a mechanism involving ribosome subunit man et al., 2012; Dimitrova et al., 2009). This pathway was discovered from the observation in yeast dissociation. Here, we reconstitute ribosome-asso- that translation of a ‘‘nonstop’’ mRNA lacking a stop codon ciated ubiquitination with purified factors to define