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PMID: 19933110 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

Structural insight into nascent polypeptide chain-mediated translational stalling.

Science (New York, N.Y.) ·Vol. 326 ·No. 5958 ·2009-12-04 ·Pages 1412-5

Seidelt B, Innis CA, Wilson DN, Gartmann M, Armache JP, Villa E, Trabuco LG, Becker T, Mielke T, Schulten K, Steitz TA, Beckmann R

Abstract

Expression of the Escherichia coli tryptophanase operon depends on ribosome stalling during translation of the upstream TnaC leader peptide, a process for which interactions between the TnaC nascent chain and the ribosomal exit tunnel are critical. We determined a 5.8 angstrom-resolution cryo-electron microscopy and single-particle reconstruction of a ribosome stalled during translation of the tnaC leader gene. The nascent chain was extended within the exit tunnel, making contacts with ribosomal components at distinct sites. Upon stalling, two conserved residues within the peptidyltransferase center adopted conformations that preclude binding of release factors. We propose a model whereby interactions within the tunnel are relayed to the peptidyltransferase center to inhibit translation. Moreover, we show that nascent chains adopt distinct conformations within the ribosomal exit tunnel.

MeSH Terms
Binding Sites Cryoelectron Microscopy Escherichia coli/genetics,metabolism Escherichia coli Proteins/chemistry,genetics,metabolism,ultrastructure Gene Expression Regulation, Bacterial Image Processing, Computer-Assisted Models, Biological Models, Molecular Operon Peptidyl Transferases/metabolism Protein Biosynthesis Protein Conformation RNA-Binding Proteins/chemistry,metabolism,ultrastructure Ribosomal Proteins/chemistry,metabolism,ultrastructure Ribosomes/metabolism,ultrastructure Tryptophanase/biosynthesis,genetics
Chemicals
Escherichia coli Proteins RNA-Binding Proteins Ribosomal Proteins rplV protein, E coli tnaC protein, E coli Peptidyl Transferases Tryptophanase
Authors & Affiliations
12 authors, click to expand affiliations / ORCID
Seidelt Birgit
Gene Center and Center for Integrated Protein Science Munich (CIPSM), Department for Chemistry and Biochemistry, University of Munich, Feodor-Lynen-Strasse 25, 81377 Munich, Germany.
Innis C Axel
Wilson Daniel N
Gartmann Marco
Armache Jean-Paul
Villa Elizabeth
Trabuco Leonardo G
Becker Thomas
Mielke Thorsten
Schulten Klaus
Steitz Thomas A
Beckmann Roland
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Article Info
Journal
Science (New York, N.Y.)
Abbr.
Science
ISSN
1095-9203
DOI
10.1126/science.1177662
Published
2009-12-04
Epub
2009-00-29
Pages
1412-5
Language
English
Region
United States
NLM ID
0404511
PMCID
PMC2920484
Subset
IM
Grants
NCRR NIH HHS · P41 RR005969 · United States
Howard Hughes Medical Institute · United States
NIGMS NIH HHS · P01 GM022778 · United States
NIGMS NIH HHS · GM022778 · United States
NCRR NIH HHS · P41-RR05969 · United States
NCRR NIH HHS · P41 RR005969-19 · United States
Databases
PDB
2WWL, 2WWQ
Corrections
CommentIn
PMID: 19965743
External Links
PubMed source DOI: 10.1126/science.1177662 PMC Full Text View journal details
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