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Title: Real-time observation of signal recognition particle binding to actively translating ribosomes

The signal recognition particle (SRP) directs translating ribosome-nascent chain complexes (RNCs) that display a signal sequence to protein translocation channels in target membranes. All previous work on the initial step of the targeting reaction, when SRP binds to RNCs, used stalled and non-translating RNCs. This meant that an important dimension of the co-translational process remained unstudied. We apply single-molecule fluorescence measurements to observe directly and in real-time E. coli SRP binding to actively translating RNCs. We show at physiologically relevant SRP concentrations that SRP-RNC association and dissociation rates depend on nascent chain length and the exposure of a functional signal sequence outside the ribosome. Our results resolve a long-standing question: how can a limited, sub-stoichiometric pool of cellular SRP effectively distinguish RNCs displaying a signal sequence from those that are not? The answer is strikingly simple: as originally proposed, SRP only stably engages translating RNCs exposing a functional signal sequence.

 
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NSF-PAR ID:
10000992
Author(s) / Creator(s):
 ;  ;  ;  
Publisher / Repository:
eLife Sciences Publications, Ltd.
Date Published:
Journal Name:
eLife
Volume:
3
ISSN:
2050-084X
Format(s):
Medium: X
Sponsoring Org:
National Science Foundation
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