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Heterologous expression of Aquifex aeolicus ribosome recycling factor in Escherichia coli is dominant lethal by forming a complex that lacks functional co‐ordination for ribosome disassembly
Author(s) -
Yamami Tohru,
Ito Koichi,
Fujiwara Toshinobu,
Nakamura Yoshikazu
Publication year - 2005
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1111/j.1365-2958.2004.04387.x
Subject(s) - biology , aquifex aeolicus , ribosome , escherichia coli , heterologous , microbiology and biotechnology , heterologous expression , translation (biology) , computational biology , genetics , recombinant dna , rna , messenger rna , gene
Summary Recycling the post‐termination ribosomal complex requires the co‐ordinated effort of the ribosome, ribosome recycling factor (RRF) and elongation factor EF‐G. Although Aquifex aeolicus RRF (aaRRF) binds Escherichia coli ribosomes as efficiently as E. coli RRF, the resulting complex is non‐functional and dominant lethal in E. coli , even in the presence of homologous A. aeolicus EF‐G. These findings suggest that the E. coli post‐termination ribosomal complex with aaRRF lacks functional co‐ordination with EF‐G required for ribosome recycling. A chimeric EF‐G ( E. coli domains I–III, A. aeolicus domains IV–V) or an A. aeolicus EF‐G with distinct mutations in the domain I–II interface could activate aaRRF. Furthermore, novel mutations that localize to one surface of the L‐shape structure of aaRRF restored activity in E. coli . These aaRRF mutations are spatially distinct from mutations previously described and suggest a novel active centre for coupling EF‐G's G domain motor action to ribosome disassembly.