Abstract
RRF (ribosome recycling factor) consists of two domains, and in concert with EF-G (elongation factor-G), triggers dissociation of the post-termination ribosomal complex. However, the function of the individual domains of RRF remains unclear. To clarify this, two RRF chimaeras, EcoDI/TteDII and TteDI/EcoDII, were created by domain swaps between the proteins from Escherichia coli and Thermoanaerobacter tengcongensis. The ribosome recycling activity of the RRF chimaeras was compared with their wild-type RRFs by using in vivo and in vitro activity assays. Like wild-type TteRRF (T. tengcongensis RRF), the EcoDI/TteDII chimaera is non-functional in E. coli, but both wild-type TYeRRF, and EcoDI/TteDII can be activated by coexpression of T. tengcongensis EF-G in E. coli. By contrast, like wild-type E. coli RRF (EcoRRF), TteDI/EcoDII is fully functional in E. coli. These findings suggest that domain II of RRF plays a crucial role in the conceited action of RRF and EF-G for the post-termination complex disassembly, and the specific interaction between RRF and EF-G on ribosomes mainly depends on the interaction between domain II of RRF and EF-G. This study provides direct genetic and biochemical evidence for the function of the individual domains of RRF.
Original language | English (US) |
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Pages (from-to) | 767-777 |
Number of pages | 11 |
Journal | Biochemical Journal |
Volume | 393 |
Issue number | 3 |
DOIs | |
State | Published - Feb 1 2006 |
Externally published | Yes |
Keywords
- Domain function
- Domain swaps
- Elongation factor (EF-G)
- Escherichia coli
- Ribosome recycling factor (RRF)
- Thermoanaerobacter tengcongensis
ASJC Scopus subject areas
- Biochemistry
- Molecular Biology
- Cell Biology