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Title: Direct measurement of the mechanical work during translocation by the ribosome

Abstract

A detailed understanding of tRNA/mRNA translocation requires measurement of the forces generated by the ribosome during this movement. Such measurements have so far remained elusive and, thus, little is known about the relation between force and translocation and how this reflects on its mechanism and regulation. Here, we address these questions using optical tweezers to follow translation by individual ribosomes along single mRNA molecules, against an applied force. We find that translocation rates depend exponentially on the force, with a characteristic distance close to the one-codon step, ruling out the existence of sub-steps and showing that the ribosome likely functions as a Brownian ratchet. We show that the ribosome generates ∼13 pN of force, barely sufficient to unwind the most stable structures in mRNAs, thus providing a basis for their regulatory role. Our assay opens the way to characterizing the ribosome's full mechano–chemical cycle.

Authors:
; ; ; ; ; ; ; ; ; ;
Publication Date:
Sponsoring Org.:
USDOE
OSTI Identifier:
1197671
Alternate Identifier(s):
OSTI ID: 1197672
Grant/Contract Number:  
AC0376SF00098(MSD KC261)
Resource Type:
Journal Article: Published Article
Journal Name:
eLife
Additional Journal Information:
Journal Name: eLife Journal Volume: 3; Journal ID: ISSN 2050-084X
Publisher:
eLife Sciences Publications, Ltd.
Country of Publication:
United States
Language:
English

Citation Formats

Liu, Tingting, Kaplan, Ariel, Alexander, Lisa, Yan, Shannon, Wen, Jin-Der, Lancaster, Laura, Wickersham, Charles E., Fredrik, Kurt, Noller, Harry, Tinoco, Ignacio, and Bustamante, Carlos J. Direct measurement of the mechanical work during translocation by the ribosome. United States: N. p., 2014. Web. doi:10.7554/eLife.03406.
Liu, Tingting, Kaplan, Ariel, Alexander, Lisa, Yan, Shannon, Wen, Jin-Der, Lancaster, Laura, Wickersham, Charles E., Fredrik, Kurt, Noller, Harry, Tinoco, Ignacio, & Bustamante, Carlos J. Direct measurement of the mechanical work during translocation by the ribosome. United States. doi:10.7554/eLife.03406.
Liu, Tingting, Kaplan, Ariel, Alexander, Lisa, Yan, Shannon, Wen, Jin-Der, Lancaster, Laura, Wickersham, Charles E., Fredrik, Kurt, Noller, Harry, Tinoco, Ignacio, and Bustamante, Carlos J. Mon . "Direct measurement of the mechanical work during translocation by the ribosome". United States. doi:10.7554/eLife.03406.
@article{osti_1197671,
title = {Direct measurement of the mechanical work during translocation by the ribosome},
author = {Liu, Tingting and Kaplan, Ariel and Alexander, Lisa and Yan, Shannon and Wen, Jin-Der and Lancaster, Laura and Wickersham, Charles E. and Fredrik, Kurt and Noller, Harry and Tinoco, Ignacio and Bustamante, Carlos J.},
abstractNote = {A detailed understanding of tRNA/mRNA translocation requires measurement of the forces generated by the ribosome during this movement. Such measurements have so far remained elusive and, thus, little is known about the relation between force and translocation and how this reflects on its mechanism and regulation. Here, we address these questions using optical tweezers to follow translation by individual ribosomes along single mRNA molecules, against an applied force. We find that translocation rates depend exponentially on the force, with a characteristic distance close to the one-codon step, ruling out the existence of sub-steps and showing that the ribosome likely functions as a Brownian ratchet. We show that the ribosome generates ∼13 pN of force, barely sufficient to unwind the most stable structures in mRNAs, thus providing a basis for their regulatory role. Our assay opens the way to characterizing the ribosome's full mechano–chemical cycle.},
doi = {10.7554/eLife.03406},
journal = {eLife},
number = ,
volume = 3,
place = {United States},
year = {Mon Aug 11 00:00:00 EDT 2014},
month = {Mon Aug 11 00:00:00 EDT 2014}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record at 10.7554/eLife.03406

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Cited by: 20 works
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Works referenced in this record:

Spontaneous Intersubunit Rotation in Single Ribosomes
journal, June 2008


Spontaneous formation of the unlocked state of the ribosome is a multistep process
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Head swivel on the ribosome facilitates translocation by means of intra-subunit tRNA hybrid sites
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