Coordinated conformational and compositional dynamics drive ribosome translocation

Jin Chen, Alexey Petrov, Albert Tsai, Seán E. O'Leary, Joseph D. Puglisi

Research output: Contribution to journalArticlepeer-review

89 Scopus citations

Abstract

During translation elongation, the ribosome compositional factors elongation factor G (EF-G; encoded by fusA) and tRNA alternately bind to the ribosome to direct protein synthesis and regulate the conformation of the ribosome. Here, we use single-molecule fluorescence with zero-mode waveguides to directly correlate ribosome conformation and composition during multiple rounds of elongation at high factor concentrations in Escherichia coli. Our results show that EF-G bound to GTP (EF-G-GTP) continuously samples both rotational states of the ribosome, binding with higher affinity to the rotated state. Upon successful accommodation into the rotated ribosome, the EF-G-ribosome complex evolves through several rate-limiting conformational changes and the hydrolysis of GTP, which results in a transition back to the nonrotated state and in turn drives translocation and facilitates release of both EF-G-GDP and E-site tRNA. These experiments highlight the power of tracking single-molecule conformation and composition simultaneously in real time.

Original languageEnglish (US)
Pages (from-to)718-727
Number of pages10
JournalNature Structural and Molecular Biology
Volume20
Issue number6
DOIs
StatePublished - Jun 2013
Externally publishedYes

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

Fingerprint Dive into the research topics of 'Coordinated conformational and compositional dynamics drive ribosome translocation'. Together they form a unique fingerprint.

Cite this