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tmRNA-SmpB complex mimics native aminoacyl-tRNAs in the A site of stalled ribosomes

Cheng, Kimberley ; Ivanova, Natalia ; Scheres, Sjors H. W. ; Pavlov, Michael Y. ; Maria Carazo, Jose ; Hebert, Hans ; Ehrenberg, Mans and Lindahl, Martin LU (2010) In Journal of Structural Biology 169(3). p.342-348
Abstract
Bacterial ribosomes stalled on faulty, often truncated, mRNAs lacking stop codons are rescued by trans-translation. It relies on an RNA molecule (tmRNA) capable of replacing the faulty mRNA with its own open reading frame (ORF). Translation of tmRNA ORF results in the tagging of faulty protein for degradation and its release from the ribosome. We used single-particle cryo-electron microscopy to visualize tmRNA together with its helper protein SmpB on the 70S Escherichia coli ribosome in states subsequent to GTP hydrolysis on elongation factor Tu (EF-Tu). Three-dimensional reconstruction and heterogeneity analysis resulted in a 15 A resolution structure of the tmRNA-SmpB complex accommodated in the A site of the ribosome, which shows that... (More)
Bacterial ribosomes stalled on faulty, often truncated, mRNAs lacking stop codons are rescued by trans-translation. It relies on an RNA molecule (tmRNA) capable of replacing the faulty mRNA with its own open reading frame (ORF). Translation of tmRNA ORF results in the tagging of faulty protein for degradation and its release from the ribosome. We used single-particle cryo-electron microscopy to visualize tmRNA together with its helper protein SmpB on the 70S Escherichia coli ribosome in states subsequent to GTP hydrolysis on elongation factor Tu (EF-Tu). Three-dimensional reconstruction and heterogeneity analysis resulted in a 15 A resolution structure of the tmRNA-SmpB complex accommodated in the A site of the ribosome, which shows that SmpB mimics the anticodon- and D-stem of native tRNAs missing in the tRNA-like domain of tmRNA. We conclude that the tmRNA-SmpB complex accommodates in the ribosomal A site very much like an aminoacyl-tRNA during protein elongation. (C) 2009 Elsevier Inc. All rights reserved. (Less)
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author
; ; ; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
tmRNA, Trans-translation, SmpB, Cryo-electron microscopy, Single-particle, Heterogeneity analysis
in
Journal of Structural Biology
volume
169
issue
3
pages
342 - 348
publisher
Elsevier
external identifiers
  • wos:000275296100010
  • scopus:76749086986
ISSN
1095-8657
DOI
10.1016/j.jsb.2009.10.015
language
English
LU publication?
yes
id
a9f69df1-9e38-4c3a-b278-8db826064bb3 (old id 1589488)
date added to LUP
2016-04-01 13:47:06
date last changed
2022-02-19 07:24:27
@article{a9f69df1-9e38-4c3a-b278-8db826064bb3,
  abstract     = {{Bacterial ribosomes stalled on faulty, often truncated, mRNAs lacking stop codons are rescued by trans-translation. It relies on an RNA molecule (tmRNA) capable of replacing the faulty mRNA with its own open reading frame (ORF). Translation of tmRNA ORF results in the tagging of faulty protein for degradation and its release from the ribosome. We used single-particle cryo-electron microscopy to visualize tmRNA together with its helper protein SmpB on the 70S Escherichia coli ribosome in states subsequent to GTP hydrolysis on elongation factor Tu (EF-Tu). Three-dimensional reconstruction and heterogeneity analysis resulted in a 15 A resolution structure of the tmRNA-SmpB complex accommodated in the A site of the ribosome, which shows that SmpB mimics the anticodon- and D-stem of native tRNAs missing in the tRNA-like domain of tmRNA. We conclude that the tmRNA-SmpB complex accommodates in the ribosomal A site very much like an aminoacyl-tRNA during protein elongation. (C) 2009 Elsevier Inc. All rights reserved.}},
  author       = {{Cheng, Kimberley and Ivanova, Natalia and Scheres, Sjors H. W. and Pavlov, Michael Y. and Maria Carazo, Jose and Hebert, Hans and Ehrenberg, Mans and Lindahl, Martin}},
  issn         = {{1095-8657}},
  keywords     = {{tmRNA; Trans-translation; SmpB; Cryo-electron microscopy; Single-particle; Heterogeneity analysis}},
  language     = {{eng}},
  number       = {{3}},
  pages        = {{342--348}},
  publisher    = {{Elsevier}},
  series       = {{Journal of Structural Biology}},
  title        = {{tmRNA-SmpB complex mimics native aminoacyl-tRNAs in the A site of stalled ribosomes}},
  url          = {{http://dx.doi.org/10.1016/j.jsb.2009.10.015}},
  doi          = {{10.1016/j.jsb.2009.10.015}},
  volume       = {{169}},
  year         = {{2010}},
}