Please use this identifier to cite or link to this item: http://repositorio.unicamp.br/jspui/handle/REPOSIP/326291
Type: Artigo
Title: Structure And Function Of The Yeast Listerin (ltn1) Conserved N-terminal Domain In Binding To Stalled 60s Ribosomal Subunits
Author: Doamekpor
Selom K.; Lee
Joong-Won; Hepowit
Nathaniel L.; Wu
Cheng; Charenton
Clement; Leonard
Marilyn; Bengtson
Mario H.; Rajashankar
Kanagalaghatta R.; Sachs
Matthew S.; Lima
Christopher D.; Joazeiro
Claudio A. P.
Abstract: 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-angstrom resolution. The structure, combined with additional mutational studies, provides insight to NTD's role in binding stalled 60S subunits. 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.
Subject: Listerin
Ltn1
Rqc
Ribosome
Structure
Editor: Natl Acad Sciences
Washington
Rights: aberto
Identifier DOI: 10.1073/pnas.1605951113
Address: http://www.pnas.org.ez88.periodicos.capes.gov.br/content/113/29/E4151.short
Date Issue: 2016
Appears in Collections:Unicamp - Artigos e Outros Documentos

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