{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T12:21:03Z","timestamp":1790857263704,"version":"4.1.0"},"reference-count":34,"publisher":"Rockefeller University Press","issue":"6","license":[{"start":{"date-parts":[[2025,4,21]],"date-time":"2025-04-21T00:00:00Z","timestamp":1745193600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/rupress.org\/pages\/terms102024"}],"funder":[{"DOI":"10.13039\/100000062","name":"National Institute of Diabetes and Digestive and Kidney Diseases","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100000062","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["R35GM145249"],"award-info":[{"award-number":["R35GM145249"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["rupress.org"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2025,6,2]]},"abstract":"<jats:p>Ribosome stalling during co-translational translocation at the ER causes translocon clogging and impairs ER protein biogenesis. Mammalian cells resolve translocon clogging via a poorly characterized translocation-associated quality control (TAQC) process. Here, we combine a genome-wide CRISPR screen with live-cell imaging to dissect the molecular linchpin of TAQC. We show that TAQC substrates translated from mRNAs bearing a ribosome-stalling poly(A) sequence are degraded by lysosomes and the proteasome. By contrast, the degradation of defective nascent chains encoded by nonstop (NS) mRNAs involves an unconventional ER-associated protein degradation (ERAD) mechanism depending on ER-to-Golgi trafficking, KDEL-mediated substrate retrieval at the Golgi, and a tRNA-binding factor NEMF that appends an aggregation-prone carboxyl tail to stalled NS nascent chains. We propose that NEMF-mediated CAT tailing targets a subset of TAQC substrates via Golgi retrieval for ERAD, safeguarding ER homeostasis.<\/jats:p>","DOI":"10.1083\/jcb.202408199","type":"journal-article","created":{"date-parts":[[2025,3,21]],"date-time":"2025-03-21T02:21:13Z","timestamp":1742523673000},"update-policy":"https:\/\/doi.org\/10.1083\/jcb.crossmarkpolicy","source":"Crossref","is-referenced-by-count":6,"title":["NEMF-mediated CAT tailing facilitates translocation-associated quality control"],"prefix":"10.1083","volume":"224","author":[{"ORCID":"https:\/\/orcid.org\/0009-0003-8694-022X","authenticated-orcid":true,"given":"Amanda","family":"Ennis","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}],"name":"National Institute of Diabetes, Digestive, and Kidney Diseases, National Institutes of Health 1 Laboratory of Molecular Biology, , Bethesda, MD,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-2896-9380","authenticated-orcid":true,"given":"Lihui","family":"Wang","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}],"name":"National Institute of Diabetes, Digestive, and Kidney Diseases, National Institutes of Health 1 Laboratory of Molecular Biology, , Bethesda, MD,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0958-8846","authenticated-orcid":true,"given":"Yue","family":"Xu","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}],"name":"National Institute of Diabetes, Digestive, and Kidney Diseases, National Institutes of Health 1 Laboratory of Molecular Biology, , Bethesda, MD,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-2768-0988","authenticated-orcid":true,"given":"Layla","family":"Saidi","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}],"name":"National Institute of Diabetes, Digestive, and Kidney Diseases, National Institutes of Health 1 Laboratory of Molecular Biology, , Bethesda, MD,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0007-6700-1277","authenticated-orcid":true,"given":"Xiaorong","family":"Wang","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/04gyf1771","id-type":"ROR","asserted-by":"publisher"}],"name":"University of California Irvine 2 Department of Physiology and Biophysics, , Irvine, CA,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2931-5474","authenticated-orcid":true,"given":"Clinton","family":"Yu","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/04gyf1771","id-type":"ROR","asserted-by":"publisher"}],"name":"University of California Irvine 2 Department of Physiology and Biophysics, , Irvine, CA,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1948-3690","authenticated-orcid":true,"given":"Sijung","family":"Yun","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}],"name":"National Institute of Diabetes, Digestive, and Kidney Diseases, National Institutes of Health 1 Laboratory of Molecular Biology, , Bethesda, MD,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3140-4687","authenticated-orcid":true,"given":"Lan","family":"Huang","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/04gyf1771","id-type":"ROR","asserted-by":"publisher"}],"name":"University of California Irvine 2 Department of Physiology and Biophysics, , Irvine, CA,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9512-7922","authenticated-orcid":true,"given":"Yihong","family":"Ye","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}],"name":"National Institute of Diabetes, Digestive, and Kidney Diseases, National Institutes of Health 1 Laboratory of Molecular Biology, , Bethesda, MD,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"291","published-online":{"date-parts":[[2025,4,22]]},"reference":[{"key":"2025042320510930800_bib1","doi-asserted-by":"crossref","first-page":"30795","DOI":"10.1038\/srep30795","article-title":"Quality control of nonstop membrane proteins at the ER membrane and in the cytosol","volume":"6","author":"Arakawa","year":"2016","journal-title":"Sci. Rep."},{"key":"2025042320510930800_bib2","doi-asserted-by":"publisher","first-page":"759","DOI":"10.1038\/nrm2999","article-title":"TRAPP complexes in membrane traffic: Convergence through a common Rab","volume":"11","author":"Barrowman","year":"2010","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"2025042320510930800_bib3","doi-asserted-by":"publisher","first-page":"jcs232850","DOI":"10.1242\/jcs.232850","article-title":"ER-associated degradation in health and disease - from substrate to organism","volume":"132","author":"Bhattacharya","year":"2019","journal-title":"J. Cell Sci."},{"key":"2025042320510930800_bib4","doi-asserted-by":"publisher","first-page":"325","DOI":"10.1038\/nsmb.2793","article-title":"Cleaning up in the endoplasmic reticulum: Ubiquitin in charge","volume":"21","author":"Christianson","year":"2014","journal-title":"Nat. Struct. Mol. Biol."},{"key":"2025042320510930800_bib5","doi-asserted-by":"publisher","first-page":"18454","DOI":"10.1074\/jbc.M115.663559","article-title":"Rkr1\/Ltn1 ubiquitin ligase-mediated degradation of translationally stalled endoplasmic reticulum proteins","volume":"290","author":"Crowder","year":"2015","journal-title":"J. Biol. Chem."},{"key":"2025042320510930800_bib6","doi-asserted-by":"publisher","first-page":"10343","DOI":"10.1074\/jbc.M808840200","article-title":"Nascent peptide-dependent translation arrest leads to Not4p-mediated protein degradation by the proteasome","volume":"284","author":"Dimitrova","year":"2009","journal-title":"J. Biol. Chem."},{"key":"2025042320510930800_bib7","doi-asserted-by":"publisher","first-page":"1451","DOI":"10.1016\/j.molcel.2022.03.038","article-title":"Ribosome-associated quality-control mechanisms from bacteria to humans","volume":"82","author":"Filbeck","year":"2022","journal-title":"Mol. Cell"},{"key":"2025042320510930800_bib8","doi-asserted-by":"publisher","first-page":"603","DOI":"10.1080\/10409238.2021.1938507","article-title":"Ribosome-associated quality control and CAT tailing","volume":"56","author":"Howard","year":"2021","journal-title":"Crit. Rev. Biochem. Mol. Biol."},{"key":"2025042320510930800_bib9","doi-asserted-by":"publisher","first-page":"1084","DOI":"10.1093\/nar\/gkz1201","article-title":"Quality controls induced by aberrant translation","volume":"48","author":"Inada","year":"2020","journal-title":"Nucleic Acids Res."},{"key":"2025042320510930800_bib10","doi-asserted-by":"publisher","first-page":"368","DOI":"10.1038\/s41580-019-0118-2","article-title":"Mechanisms and functions of ribosome-associated protein quality control","volume":"20","author":"Joazeiro","year":"2019","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"2025042320510930800_bib11","doi-asserted-by":"publisher","first-page":"743","DOI":"10.1016\/j.molcel.2016.11.039","article-title":"Initiation of quality control during Poly(A) translation requires site-specific ribosome ubiquitination","volume":"65","author":"Juszkiewicz","year":"2017","journal-title":"Mol. Cell"},{"key":"2025042320510930800_bib12","doi-asserted-by":"publisher","first-page":"414","DOI":"10.1126\/science.aam7787","article-title":"CAT-tailing as a fail-safe mechanism for efficient degradation of stalled nascent polypeptides","volume":"357","author":"Kostova","year":"2017","journal-title":"Science"},{"key":"2025042320510930800_bib13","doi-asserted-by":"publisher","first-page":"113860","DOI":"10.1016\/j.celrep.2024.113860","article-title":"NEMF-mediated Listerin-independent mitochondrial translational surveillance by E3 ligase Pirh2 and mitochondrial protease ClpXP","volume":"43","author":"Lv","year":"2024","journal-title":"Cell Rep."},{"key":"2025042320510930800_bib14","doi-asserted-by":"publisher","first-page":"15981","DOI":"10.1073\/pnas.1413882111","article-title":"Structural basis for translational surveillance by the large ribosomal subunit-associated protein quality control complex","volume":"111","author":"Lyumkis","year":"2014","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"2025042320510930800_bib15","doi-asserted-by":"publisher","first-page":"10","DOI":"10.14806\/ej.17.1.200","article-title":"Cutadapt removes adapter sequences from high-throughput sequencing reads","volume":"17","author":"Martin","year":"2011","journal-title":"EMBnet J."},{"key":"2025042320510930800_bib16","doi-asserted-by":"publisher","first-page":"314","DOI":"10.1080\/19491034.2015.1079680","article-title":"Principles of protein targeting to the nucleolus","volume":"6","author":"Martin","year":"2015","journal-title":"Nucleus"},{"key":"2025042320510930800_bib17","doi-asserted-by":"publisher","first-page":"2102","DOI":"10.1093\/nar\/gkab005","article-title":"The nascent polypeptide in the 60S subunit determines the Rqc2-dependency of ribosomal quality control","volume":"49","author":"Mizuno","year":"2021","journal-title":"Nucleic Acids Res."},{"key":"2025042320510930800_bib18","doi-asserted-by":"publisher","first-page":"869","DOI":"10.1083\/jcb.201312042","article-title":"Quality control: ER-associated degradation: Protein quality control and beyond","volume":"204","author":"Ruggiano","year":"2014","journal-title":"J. Cell Biol."},{"key":"2025042320510930800_bib19","doi-asserted-by":"publisher","first-page":"783","DOI":"10.1038\/nmeth.3047","article-title":"Improved vectors and genome-wide libraries for CRISPR screening","volume":"11","author":"Sanjana","year":"2014","journal-title":"Nat. Methods"},{"key":"2025042320510930800_bib20","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2220340120","article-title":"RPL26\/uL24 UFMylation is essential for ribosome-associated quality control at the endoplasmic reticulum","volume":"120","author":"Scavone","year":"2023","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"2025042320510930800_bib21","doi-asserted-by":"publisher","first-page":"45","DOI":"10.1083\/jcb.147.1.45","article-title":"The pathway of US11-dependent degradation of MHC class I heavy chains involves a ubiquitin-conjugated intermediate","volume":"147","author":"Shamu","year":"1999","journal-title":"J. Cell Biol."},{"key":"2025042320510930800_bib22","doi-asserted-by":"publisher","first-page":"433","DOI":"10.1016\/j.molcel.2014.12.015","article-title":"Structure and assembly pathway of the ribosome quality control complex","volume":"57","author":"Shao","year":"2015","journal-title":"Mol. Cell"},{"key":"2025042320510930800_bib23","doi-asserted-by":"publisher","first-page":"75","DOI":"10.1126\/science.1259724","article-title":"Protein synthesis. Rqc2p and 60S ribosomal subunits mediate mRNA-independent elongation of nascent chains","volume":"347","author":"Shen","year":"2015","journal-title":"Science"},{"key":"2025042320510930800_bib24","doi-asserted-by":"publisher","first-page":"450","DOI":"10.1038\/s41594-019-0230-1","article-title":"CAT tails drive degradation of stalled polypeptides on and off the ribosome","volume":"26","author":"Sitron","year":"2019","journal-title":"Nat. Struct. Mol. Biol."},{"key":"2025042320510930800_bib25","doi-asserted-by":"publisher","first-page":"607","DOI":"10.1016\/j.molcel.2023.01.020","article-title":"Molecular basis of eIF5A-dependent CAT tailing in eukaryotic ribosome-associated quality control","volume":"83","author":"Tesina","year":"2023","journal-title":"Mol. Cell"},{"key":"2025042320510930800_bib26","doi-asserted-by":"publisher","first-page":"2112","DOI":"10.1016\/j.molcel.2021.03.004","article-title":"Convergence of mammalian RQC and C-end rule proteolytic pathways via alanine tailing","volume":"81","author":"Thrun","year":"2021","journal-title":"Mol. Cell"},{"key":"2025042320510930800_bib27","doi-asserted-by":"publisher","first-page":"944","DOI":"10.1038\/nrm2546","article-title":"One step at a time: Endoplasmic reticulum-associated degradation","volume":"9","author":"Vembar","year":"2008","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"2025042320510930800_bib28","doi-asserted-by":"publisher","first-page":"1299","DOI":"10.1073\/pnas.1816202116","article-title":"Ribosomal protein RPL26 is the principal target of UFMylation","volume":"116","author":"Walczak","year":"2019","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"2025042320510930800_bib29","doi-asserted-by":"publisher","first-page":"5","DOI":"10.1038\/s41422-019-0236-6","article-title":"UFMylation of RPL26 links translocation-associated quality control to endoplasmic reticulum protein homeostasis","volume":"30","author":"Wang","year":"2020","journal-title":"Cell Res."},{"key":"2025042320510930800_bib30","doi-asserted-by":"publisher","first-page":"112028","DOI":"10.1016\/j.celrep.2023.112028","article-title":"SAYSD1 senses UFMylated ribosome to safeguard co-translational protein translocation at the endoplasmic reticulum","volume":"42","author":"Wang","year":"2023","journal-title":"Cell Rep."},{"key":"2025042320510930800_bib31","doi-asserted-by":"publisher","first-page":"610689","DOI":"10.3389\/fcell.2020.610689","article-title":"Clearing traffic jams during protein translocation across membranes","volume":"8","author":"Wang","year":"2021","journal-title":"Front. Cell Dev. Biol."},{"key":"2025042320510930800_bib32","doi-asserted-by":"publisher","first-page":"652","DOI":"10.1038\/414652a","article-title":"The AAA ATPase Cdc48\/p97 and its partners transport proteins from the ER into the cytosol","volume":"414","author":"Ye","year":"2001","journal-title":"Nature"},{"key":"2025042320510930800_bib33","doi-asserted-by":"publisher","first-page":"731","DOI":"10.1016\/j.tibs.2021.03.008","article-title":"Detecting and rescuing stalled ribosomes","volume":"46","author":"Yip","year":"2021","journal-title":"Trends Biochem. Sci."},{"key":"2025042320510930800_bib34","doi-asserted-by":"publisher","first-page":"4438","DOI":"10.1091\/mbc.E15-06-0354","article-title":"gp78 functions downstream of Hrd1 to promote degradation of misfolded proteins of the endoplasmic reticulum","volume":"26","author":"Zhang","year":"2015","journal-title":"Mol. Biol. Cell"}],"container-title":["Journal of Cell Biology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/rupress.org\/jcb\/article-pdf\/doi\/10.1083\/jcb.202408199\/1942792\/jcb_202408199.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/rupress.org\/jcb\/article-pdf\/doi\/10.1083\/jcb.202408199\/1942792\/jcb_202408199.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,4,23]],"date-time":"2025-04-23T20:51:19Z","timestamp":1745441479000},"score":1,"resource":{"primary":{"URL":"https:\/\/rupress.org\/jcb\/article\/224\/6\/e202408199\/277400\/NEMF-mediated-CAT-tailing-facilitates"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,4,22]]},"references-count":34,"journal-issue":{"issue":"6","published-print":{"date-parts":[[2025,6,2]]}},"URL":"https:\/\/doi.org\/10.1083\/jcb.202408199","relation":{},"ISSN":["0021-9525","1540-8140"],"issn-type":[{"value":"0021-9525","type":"print"},{"value":"1540-8140","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,4,22]]},"article-number":"e202408199"}}