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Volume 148, Issue 12

Characterization of ribosome recycling factor (RRF) and a mutant lacking six amino acids from the C-terminal end reveals that the C-terminal residues are important for its occupancy on the ribosome Free

Abstract

Ribosome recycling factor (RRF), coded for by the locus, is involved in the disassembly of post-termination complexes and recycling of the ribosomes for a fresh round of initiation in bacteria and in eukaryotic organelles. In a cross-species-complementation experiment, it was shown that the RRF protein lacking five amino acids from its C-terminal end (ΔC5RRF) but not the full-length protein (RRF) complemented for its phenotype. It was also shown that the RFF protein (RRF) did not complement LJ14 for . However, simultaneous expression of elongation factor G (EFG) and RRF from resulted in complementation of LJ14. Here it is shown that unlike ΔC5RRF, an equivalent mutant of RRF lacking six amino acids from its C-terminal end (ΔC6RRF) did not complement LJ14. Surprisingly, ΔC6RRF failed to complement the strain even in the presence of homologous EFG (EFG). The biochemical and biophysical characterization of these proteins suggested that the mutant RRF folded properly. However, ribosome-binding assays showed that the mutant protein was compromised in its binding to ribosomes. It is suggested that the conserved amino acids at the C-terminal end of the RRFs contribute to their residency on ribosomes and that the specific interactions between RRF and EFG are crucial in the disassembly of the termination complex.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
Keyword(s): CD, circular dichroism , EcoEFG, E. coli EFG , EcoRRF, full-length E. coli RRF , EFG, elongation factor G; frr, locus coding for RRF , elongation factor G , ESI-MS, electron spray ionization-mass spectroscopy , frrts, locus coding for temperature-sensitive RRF , MtuEFG, M. tuberculosis EFG , MtuRRF, full-length M. tuberculosis RRF , polysome binding , RFF, ribosome recycling factor , termination complex , Thermus thermophilus , TthRRF, full-length T. thermophilus RRF and tubercle bacilli
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2002-12-01
2025-01-22
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/content/journal/micro/10.1099/00221287-148-12-3913
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Characterization of Mycobacterium tuberculosis ribosome recycling factor (RRF) and a mutant lacking six amino acids from the C-terminal end reveals that the C-terminal residues are important for its occupancy on the ribosome
Microbiology 148, 3913 (2002); https://doi.org/10.1099/00221287-148-12-3913
/content/journal/micro/10.1099/00221287-148-12-3913
/content/journal/micro/10.1099/00221287-148-12-3913
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