RELRCSBRCSBRCSB2014-11-042014-12-242015-01-072015-01-21Electron cryo-microscopy of Ltn1, Rqc2 bound to yeast 60S ribosomeShen PSPark JQin YLi XParsawar KLarson MCox JCheng YLambowitz AMWeissman JSBrandman OFrost Aribosome quality control complex, RQC, eukaryotic ribosome rescue, stalled nascent chainShen PSPark JQin YLi XParsawar KLarson MHCox JCheng YLambowitz AMWeissman JSBrandman OFrost ARqc2p and 60S ribosomal subunits mediate mRNA-independent elongation of nascent chainsSCIENCE3477578201525554787doi:10.1126/science.1259724RQC particles purified by co-IP of Rqc1-FLAG, eluted with 3xFLAG peptideRQC particles purified by co-IP of Rqc1-FLAG, eluted with 3xFLAG peptideRQC particles were 3D classified to reveal distinct subclasses containing various RQC components.160S ribosomefalseSaccharomyces cerevisiaeBY4741yeastribosome-eukaryote: LSU 60S, LSU RNA 28S, LSU RNA 5.8S, LSU RNA 5SsingleParticleparticle7.4100 mM KOAc, 10 mM MgCl2, 25 mM HEPES-KOH200 mesh Quantifoil R2/2 grid + lacey carbon grid with ultrathin carbonETHANE7590FEI VITROBOT MARK IIIBlot for 3 seconds before plungingFEI POLARA 300FLOOD BEAMBRIGHT FIELDFIELD EMISSION GUN3002.00.82.031000.0GATAN LIQUID NITROGEN809085GatanUCSF Image4 on-the-fly motion correction2013-07-22GATAN K2 (4k x 4k)3459358.LN2 cooledParticles were selected using the semi-automated swarm tool in e2boxer.py of the EMAN2 package. All 2D and 3D processing was performed in RELION.each particle8.2OTHERRELION, CTFFIND3Micrographs were motion-corrected via the UCSFImage4 package.9131