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Activated GTPase movement on an RNA scaffold drives co-translational protein targeting

Shen, Kuang and Arslan, Sinan and Akopian, David and Ha, Taekjip and Shan, Shu-ou (2012) Activated GTPase movement on an RNA scaffold drives co-translational protein targeting. Nature, 492 (7428). pp. 271-275. ISSN 0028-0836. PMCID PMC3531814. doi:10.1038/nature11726. https://resolver.caltech.edu/CaltechAUTHORS:20130103-133200806

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Abstract

Approximately one-third of the proteome is initially destined for the eukaryotic endoplasmic reticulum or the bacterial plasma membrane. The proper localization of these proteins is mediated by a universally conserved protein-targeting machinery, the signal recognition particle (SRP), which recognizes ribosomes carrying signal sequences and, through interactions with the SRP receptor delivers them to the protein-translocation machinery on the target membrane. The SRP is an ancient ribonucleoprotein particle containing an essential, elongated SRP RNA for which precise functions have remained elusive. Here we used single-molecule fluorescence microscopy to show that the Escherichia coli SRP–SRP receptor GTPase complex, after initial assembly at the tetraloop end of SRP RNA, travels over 100 Å to the distal end of this RNA, where rapid GTP hydrolysis occurs. This movement is negatively regulated by the translating ribosome and, at a later stage, positively regulated by the SecYEG translocon, providing an attractive mechanism for ensuring the productive exchange of the targeting and translocation machineries at the ribosome exit site with high spatial and temporal accuracy. Our results show that large RNAs can act as molecular scaffolds that enable the easy exchange of distinct factors and precise timing of molecular events in a complex cellular process; this concept may be extended to similar phenomena in other ribonucleoprotein complexes.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1038/nature11726DOIArticle
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3531814/PubMed CentralArticle
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ORCID:
AuthorORCID
Shan, Shu-ou0000-0002-6526-1733
Additional Information:© 2012 Macmillan Publishers Limited. Received 31 July; accepted 26 October 2012. Published online 12 December 2012. We thank N. Ban and members of the Shan group for comments on the manuscript, C. Richards, L. Cai, T. Zhiyentayev, K. Lee and R. Zhou for help with RNC labelling and the instrument and software setup, and C. L. Guo, S. Kouand H. Lester for discussions. This work is supported by National Institutes of Health (NIH) grant GM078024 to S.-o.S., an NIH instrument supplement to grant GM45162 to D.C. Rees, and Caltech matching fund 350270 for the single-molecule instruments. S.-o.S. was supported by the Beckman Young Investigator award, the David and Lucile Packard Fellowship in Science and Engineering, and the Henry Dreyfus Teacher-Scholar award. T. H. was supported by National Science FoundationPhysics Frontiers Centers program (08222613) and NIH grant GM065367. Author Contributions: K.S., S.A., T.H. and S.-o.S. conceived the experiments. K.S. purified and labelled Ffh, FtsY, DNA, RNA and RNC. D.A. purified SecYEG and performed the GTPase assay in Fig. 4a. K.S. and S.A. carried out smFRET measurements under the direction of T.H. K.S. and S.A. analysed the data. K.S. and S.-o.S. wrote the paper with inputs from all other authors.
Funders:
Funding AgencyGrant Number
NIHGM078024
NIHGM45162
Caltech Matching Fund350270
Arnold and Mabel Beckman FoundationUNSPECIFIED
David and Lucile Packard FoundationUNSPECIFIED
Camille and Henry Dreyfus FoundationUNSPECIFIED
NSFPHY-08222613
NIHGM065367
Subject Keywords:Enzyme mechanisms; Protein translocation; Single-molecule biophysics; RNA
Issue or Number:7428
PubMed Central ID:PMC3531814
DOI:10.1038/nature11726
Record Number:CaltechAUTHORS:20130103-133200806
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20130103-133200806
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:36150
Collection:CaltechAUTHORS
Deposited By: Jason Perez
Deposited On:04 Jan 2013 15:42
Last Modified:09 Nov 2021 23:20

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