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Ribosome Binding of a Single Copy of the SecY Complex: Implications for Protein Translocation

Ménétret, Jean-François and Schaletzky, Julia and Clemons, William M., Jr. and Osborne, Andrew R. and Skånland, Sigrid S. and Denison, Carilee and Gygi, Steven P. and Kirkpatrick, Don S. and Park, Eunyong and Ludtke, Steven J. and Rapoport, Tom A. and Akey, Christopher W. (2007) Ribosome Binding of a Single Copy of the SecY Complex: Implications for Protein Translocation. Molecular Cell, 28 (6). pp. 1083-1092. ISSN 1097-2765. https://resolver.caltech.edu/CaltechAUTHORS:20181101-114159777

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Abstract

The SecY complex associates with the ribosome to form a protein translocation channel in the bacterial plasma membrane. We have used cryo-electron microscopy and quantitative mass spectrometry to show that a nontranslating E. coli ribosome binds to a single SecY complex. The crystal structure of an archaeal SecY complex was then docked into the electron density maps. In the resulting model, two cytoplasmic loops of SecY extend into the exit tunnel near proteins L23, L29, and L24. The loop between transmembrane helices 8 and 9 interacts with helices H59 and H50 in the large subunit RNA, while the 6/7 loop interacts with H7. We also show that point mutations of basic residues within either loop abolish ribosome binding. We suggest that SecY binds to this primary site on the ribosome and subsequently captures and translocates the nascent chain.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1016/j.molcel.2007.10.034DOIArticle
ORCID:
AuthorORCID
Clemons, William M., Jr.0000-0002-0021-889X
Additional Information:© 2007 Elsevier. Under an Elsevier user license. Received 26 April 2007, Revised 3 August 2007, Accepted 4 October 2007, Available online 27 December 2007. We thank J. Rush (Cell Signaling Technology, Inc.) for synthesis of the AQUA peptides. J.S. was supported by the Boehringer Ingelheim Fonds, and W.M.C. was supported by a Damon Runyon Cancer Fellowship. Work in the T.A.R. and C.W.A. laboratories was supported by National Institutes of Health grants. T.A.R. is an investigator of the Howard Hughes Medical Institute. Accession Numbers: The modeled ribosome-SecY complexes have been deposited in the Protein Data Bank under ID codes 3BO0 and 3BO1, and the EM map has been deposited in the Electron Microscopy Database (EMD-5907).
Funders:
Funding AgencyGrant Number
Boehringer Ingelheim FondsUNSPECIFIED
Damon Runyon Cancer Research FoundationUNSPECIFIED
NIHUNSPECIFIED
Howard Hughes Medical Institute (HHMI)UNSPECIFIED
Issue or Number:6
Record Number:CaltechAUTHORS:20181101-114159777
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20181101-114159777
Official Citation:Jean-François Ménétret, Julia Schaletzky, William M. Clemons, Andrew R. Osborne, Sigrid S. Skånland, Carilee Denison, Steven P. Gygi, Don S. Kirkpatrick, Eunyong Park, Steven J. Ludtke, Tom A. Rapoport, Christopher W. Akey, Ribosome Binding of a Single Copy of the SecY Complex: Implications for Protein Translocation, Molecular Cell, Volume 28, Issue 6, 2007, Pages 1083-1092, ISSN 1097-2765, https://doi.org/10.1016/j.molcel.2007.10.034. (http://www.sciencedirect.com/science/article/pii/S1097276507008258)
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:90566
Collection:CaltechAUTHORS
Deposited By: George Porter
Deposited On:01 Nov 2018 19:54
Last Modified:03 Oct 2019 20:26

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