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Multiple conformational switches in a GTPase complex control co-translational protein targeting

Zhang, Xin and Schaffitzel, Christiane and Ban, Nenad and Shan, Shu-ou (2009) Multiple conformational switches in a GTPase complex control co-translational protein targeting. Proceedings of the National Academy of Sciences of the United States of America, 106 (6). pp. 1754-1759. ISSN 0027-8424. PMCID PMC2644110. http://resolver.caltech.edu/CaltechAUTHORS:20090407-091223238

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Abstract

The “GTPase switch” paradigm, in which a GTPase switches between an active, GTP-bound state and an inactive, GDP-bound state through the recruitment of nucleotide exchange factors (GEFs) or GTPase activating proteins (GAPs), has been used to interpret the regulatory mechanism of many GTPases. A notable exception to this paradigm is provided by two GTPases in the signal recognition particle (SRP) and the SRP receptor (SR) that control the co-translational targeting of proteins to cellular membranes. Instead of the classical “GTPase switch,” both the SRP and SR undergo a series of discrete conformational rearrangements during their interaction with one another, culminating in their reciprocal GTPase activation. Here, we show that this series of rearrangements during SRP-SR binding and activation provide important control points to drive and regulate protein targeting. Using real-time fluorescence, we showed that the cargo for SRP—ribosomes translating nascent polypeptides with signal sequences—accelerates SRP·SR complex assembly over 100-fold, thereby driving rapid delivery of cargo to the membrane. A series of subsequent rearrangements in the SRP·SR GTPase complex provide important driving forces to unload the cargo during late stages of protein targeting. Further, the cargo delays GTPase activation in the SRP·SR complex by 8–12 fold, creating an important time window that could further improve the efficiency and fidelity of protein targeting. Thus, the SRP and SR GTPases, without recruiting external regulatory factors, constitute a self-sufficient system that provides exquisite spatial and temporal control of a complex cellular process.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1073/pnas.0808573106DOIArticle
http://www.pnas.org/content/106/6/1754PublisherArticle
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2644110/PubMed CentralArticle
ORCID:
AuthorORCID
Shan, Shu-ou0000-0002-6526-1733
Additional Information:© 2009 by the National Academy of Sciences. Edited by Jennifer A. Doudna, University of California, Berkeley, CA, and approved December 22, 2008 (received for review August 28, 2008). We thank Sandra Schmid, Douglas C. Rees, Raymond Deshaies, Nathan Pierce, and members of the Shan laboratory for comments on the manuscript. This work was supported by a National Institutes of Health grant GM078024 to S.S., and by the Swiss National Science Foundation (SNSF) and the NCCR Structural Biology program of the SNSF to N.B. S.S. was supported by a career award from the Burroughs Welcome Foundation, the Henry and Camille Dreyfus foundation, the Beckman Young Investigator award, and the Packard and Lucile award in science and engineering. X.Z. was supported by a fellowship from the Ulric B. and Evelyn L. Bray Endowment Fund.
Funders:
Funding AgencyGrant Number
NIHGM078024
Swiss National Science Foundation (SNSF)UNSPECIFIED
Burroughs Welcome FoundationUNSPECIFIED
Camille and Henry Dreyfus FoundationUNSPECIFIED
Arnold and Mabel Beckman FoundationUNSPECIFIED
David and Lucile Packard FoundationUNSPECIFIED
Ulric B. and Evelyn L. Bray Endowment FundUNSPECIFIED
Subject Keywords:conformational change; fluorescence spectroscopy; protein targeting and translocation; signal recognition particle
PubMed Central ID:PMC2644110
Record Number:CaltechAUTHORS:20090407-091223238
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20090407-091223238
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:13862
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:15 Jul 2009 21:20
Last Modified:31 Jan 2017 19:39

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