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  GTP hydrolysis by EF‐G synchronizes tRNA movement on small and large ribosomal subunits.

Holtkamp, W., da Cunha, C. E. L., Peske, F., Konevega, A. L., Wintermeyer, W., & Rodnina, M. V. (2014). GTP hydrolysis by EF‐G synchronizes tRNA movement on small and large ribosomal subunits. EMBO Journal, 33(9), 1073-1085. doi:10.1002/embj.201387465.

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Holtkamp, W.1, Author           
da Cunha, C. E. L.1, Author           
Peske, F.1, Author           
Konevega, A. L.1, Author           
Wintermeyer, W.2, Author           
Rodnina, M. V.1, Author           
Affiliations:
1Department of Physical Biochemistry, MPI for biophysical chemistry, Max Planck Society, ou_578598              
2Research Group of Ribosome Dynamics, MPI for biophysical chemistry, Max Planck Society, ou_578599              

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 Abstract: Elongation factor G (EF‐G) promotes the movement of two tRNAs and the mRNA through the ribosome in each cycle of peptide elongation. During translocation, the tRNAs transiently occupy intermediate positions on both small (30S) and large (50S) ribosomal subunits. How EF‐G and GTP hydrolysis control these movements is still unclear. We used fluorescence labels that specifically monitor movements on either 30S or 50S subunits in combination with EF‐G mutants and translocation‐specific antibiotics to investigate timing and energetics of translocation. We show that EF‐G–GTP facilitates synchronous movements of peptidyl‐tRNA on the two subunits into an early post‐translocation state, which resembles a chimeric state identified by structural studies. EF‐G binding without GTP hydrolysis promotes only partial tRNA movement on the 50S subunit. However, rapid 30S translocation and the concomitant completion of 50S translocation require GTP hydrolysis and a functional domain 4 of EF‐G. Our results reveal two distinct modes for utilizing the energy of EF‐G binding and GTP hydrolysis and suggest that coupling of GTP hydrolysis to translocation is mediated through rearrangements of the 30S subunit.

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Language(s): eng - English
 Dates: 2014-03-102014-04-01
 Publication Status: Issued
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 Rev. Type: Peer
 Identifiers: DOI: 10.1002/embj.201387465
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Title: EMBO Journal
Source Genre: Journal
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Pages: - Volume / Issue: 33 (9) Sequence Number: - Start / End Page: 1073 - 1085 Identifier: -