skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Structure and function of the yeast listerin (Ltn1) conserved N-terminal domain in binding to stalled 60S ribosomal subunits

Journal Article · · Proceedings of the National Academy of Sciences of the United States of America
 [1];  [2];  [3];  [4];  [1];  [3];  [5];  [6];  [4];  [1];  [7]
  1. Sloan-Kettering Inst., New York, NY (United States)
  2. The Scripps Research Inst., La Jolla, CA (United States); The Catholic Univ. of Korea, Seoul (Korea)
  3. The Scripps Research Inst., La Jolla, CA (United States)
  4. Texas A & M Univ., College Station, TX (United States)
  5. The Scripps Research Inst., La Jolla, CA (United States); Univ. of Campinas (UNICAMP), SP (Brazil)
  6. Advanced Photon Source, Argonne, IL (United States); Cornell Univ., Ithaca, NY (United States)
  7. The Scripps Research Inst., La Jolla, CA (United States); Zentrum für Molekulare Biologie der Universität Heidelberg (Germany); Deutsches Krebsforschungszentrum–Zentrum für Molekulare Biologie der Universität Heidelberg Alliance (ZMBH-DKFZ) (Germany)
+ Show Author Affiliations

The Ltn1 E3 ligase (listerin in mammals) has emerged as a paradigm for understanding ribosome-associated ubiquitylation. Ltn1 binds to 60S ribosomal subunits to ubiquitylate nascent polypeptides that become stalled during synthesis; among Ltn1’s substrates are aberrant products of mRNA lacking stop codons [nonstop translation products (NSPs)]. Here, we report the reconstitution of NSP ubiquitylation in Neurospora crassa cell extracts. Upon translation in vitro, ribosome-stalled NSPs were ubiquitylated in an Ltn1-dependent manner, while still ribosome-associated. Furthermore, we provide biochemical evidence that the conserved N-terminal domain (NTD) plays a significant role in the binding of Ltn1 to 60S ribosomal subunits and that NTD mutations causing defective 60S binding also lead to defective NSP ubiquitylation, without affecting Ltn1’s intrinsic E3 ligase activity. Finally, we report the crystal structure of the Ltn1 NTD at 2.4-Å resolution. The structure, combined with additional mutational studies, provides insight to NTD’s role in binding stalled 60S subunits. Furthermore, our findings show that Neurospora extracts can be used as a tool to dissect mechanisms underlying ribosome-associated protein quality control and are consistent with a model in which Ltn1 uses 60S subunits as adapters, at least in part via its NTD, to target stalled NSPs for ubiquitylation.

Research Organization:
Argonne National Laboratory (ANL), Argonne, IL (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Basic Energy Sciences (BES). Scientific User Facilities Division; National Inst. of Neurological Disorders and Stroke (NINDS); National Institutes of Health (NIH); National Cancer Institute (NCI); National Inst. of General Medical Sciences; NIH-ORIP HEI
Grant/Contract Number:
AC02-06CH11357; NS075719; CA152103; GM068087; R01 GM061906; P30 CA008748; P41 GM103403; S10 RR029205
OSTI ID:
1274771
Journal Information:
Proceedings of the National Academy of Sciences of the United States of America, Vol. 113, Issue 29; ISSN 0027-8424
Publisher:
National Academy of SciencesCopyright Statement
Country of Publication:
United States
Language:
ENGLISH
Citation Metrics:
Cited by: 25 works
Citation information provided by
Web of Science

References (33)

The Use of Fungal In Vitro Systems for Studying Translational Regulation book January 2007
A Ribosome-Bound Quality Control Complex Triggers Degradation of Nascent Peptides and Signals Translation Stress journal November 2012
The Rqc2/Tae2 subunit of the ribosome-associated quality control (RQC) complex marks ribosome-stalled nascent polypeptide chains for aggregation journal March 2016
A General Procedure for the Production of Antibody Reagents Against Eukaryotic Ribosomal Proteins journal August 2005
A short history of SHELX journal December 2007
Cdc48-associated complex bound to 60S particles is required for the clearance of aberrant translation products journal March 2013
Differential Scales of Protein Quality Control journal March 2014
Ubiquitination of newly synthesized proteins at the ribosome journal July 2015
Failure of RQC machinery causes protein aggregation and proteotoxic stress journal February 2016
Coot model-building tools for molecular graphics journal November 2004
Dom34:Hbs1 Promotes Subunit Dissociation and Peptidyl-tRNA Drop-Off to Initiate No-Go Decay journal October 2010
Role of a ribosome-associated E3 ubiquitin ligase in protein quality control journal September 2010
Reconstitution of a Minimal Ribosome-Associated Ubiquitination Pathway with Purified Factors journal September 2014
Cdc48/p97 promotes degradation of aberrant nascent polypeptides bound to the ribosome journal January 2013
Electrostatics in the Ribosomal Tunnel Modulate Chain Elongation Rates journal December 2008
Structural basis for translational surveillance by the large ribosomal subunit-associated protein quality control complex journal October 2014
Dissociation by Pelota, Hbs1 and ABCE1 of mammalian vacant 80S ribosomes and stalled elongation complexes: Ribosomal dissociation by Pelota/Hbs1/ABCE1 journal March 2011
Single-particle EM reveals extensive conformational variability of the Ltn1 E3 ligase journal January 2013
A mouse forward genetics screen identifies LISTERIN as an E3 ubiquitin ligase involved in neurodegeneration journal February 2009
Ribosome-associated protein quality control journal January 2016
Translation of CGA codon repeats in yeast involves quality control components and ribosomal protein L1 journal July 2013
NMRPipe: A multidimensional spectral processing system based on UNIX pipes journal November 1995
Structure of the no-go mRNA decay complex Dom34–Hbs1 bound to a stalled 80S ribosome journal May 2011
Listerin-Dependent Nascent Protein Ubiquitination Relies on Ribosome Subunit Dissociation journal June 2013
Rqc2p and 60 S ribosomal subunits mediate mRNA-independent elongation of nascent chains journal January 2015
Polymerase Stalling during Replication, Transcription and Translation journal May 2014
Selenomethionyl proteins produced for analysis by multiwavelength anomalous diffraction (MAD): a vehicle for direct determination of three-dimensional structure. journal May 1990
Positively Charged Residues Are the Major Determinants of Ribosomal Velocity journal March 2013
Phaser crystallographic software journal July 2007
Protecting the proteome: Eukaryotic cotranslational quality control pathways journal February 2014
Structure and Assembly Pathway of the Ribosome Quality Control Complex journal February 2015
Protein Repeats: Structures, Functions, and Evolution journal May 2001
[20] Processing of X-ray diffraction data collected in oscillation mode book January 1997

Cited By (8)

CAT tails drive degradation of stalled polypeptides on and off the ribosome journal May 2019
Mechanisms and functions of ribosome-associated protein quality control journal April 2019
CAT tails drive on- and off-ribosome degradation of stalled polypeptides posted_content November 2018
Expanding Role of Ubiquitin in Translational Control journal February 2020
In vitro analysis of RQC activities provides insights into the mechanism and function of CAT tailing journal July 2017
Functional analysis of a nonstop mutation in MITF gene identified in a patient with Waardenburg syndrome type 2 journal March 2017
The ribosome-bound quality control complex remains associated to aberrant peptides during their proteasomal targeting and interacts with Tom1 to limit protein aggregation journal May 2017
Mapping the mammalian ribosome quality control complex interactome using proximity labeling approaches journal May 2018

Similar Records

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
Listerin
Ltn1
RQC
ribosome
structure