Citation: Peng Sijia, Sun Ruirui, Wang Wenjuan, Chen Chunlai. Single-molecule FRET studies on interactions between elongation factor 4 (LepA) and ribosomes[J]. Chinese Chemical Letters, ;2018, 29(10): 1503-1508. doi: 10.1016/j.cclet.2017.12.006 shu

Single-molecule FRET studies on interactions between elongation factor 4 (LepA) and ribosomes

a.
School of Life Sciences, Tsinghua-Peking Joint Center for Life Sciences, Beijing Advanced Innovation Center for Structural Biology, Tsinghua University, Beijing 100084, China
b.
School of Life Sciences and Technology Center for Protein Sciences, Tsinghua University, Beijing 100084, China

Corresponding author: Wang Wenjuan, wangwenjuan@biomed.tsinghua.edu.cn Chen Chunlai, chunlai@mail.tsinghua.edu.cn
Received Date: 2 December 2017
Revised Date: 20 December 2017
Accepted Date: 21 December 2017
Available Online: 8 October 2017

Figures(5)

Elongation factor 4 (EF4) is one of the highly conserved translational GTPases, whose functions are largely unknown. Structures of EF4 bound ribosomal PRE-translocation and POST-translocation complexes have both been visualized. On top of cellular, structural, and biochemical studies, several controversial models have been raised to rationalize functions of EF4. However, how EF4 modulates elongation through its interactions with ribosomes has not been revealed. Here, using single-molecule fluorescence resonance energy transfer assays, we directly captured short-lived EF4·GTP bound ribosomal PRE and POST translocation complexes, which may adopt slightly different conformations from structures prepared using GDP, GDPNP, or GDPCP. Furthermore, we revealed that EF4·GTP severely impairs delivery of aminoacyl-tRNA into the A-site of the ribosome and moderately accelerates translocation. We proposed that functions of EF4 are to slow overall elongation and to stall majority of ribosomes in POSTstates under stress conditions.
- Ribosome,
- Single-molecule biophysics,
- Translation,
- Fluorescence resonance energy transfer,
- GTPase
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