A dynamic competition between release factor 2 and the tRNA(Sec) decoding UGA at the recoding site of Escherichia coli formate dehydrogenase H.
Factors affecting competition between termination and elongation in vivo during translation of the fdhF selenocysteine recoding site (UGA) were studied with wild-type and modified fdhF sequences. Altering sequences surrounding the recoding site UGA without affecting RNA secondary structure indicated that the kinetics of stop signal decoding have a significant influence on selenocysteine incorporation efficiency. The UGA in the wild-type fdhF sequence remains 'visible' to the factor and forms a site-directed cross-link when mRNA stem-loop secondary structure is absent, but not when it is present. The timing of the secondary structure unfolding during translation may be a critical feature of competition between release factor 2 and tRNA(Sec) for decoding UGA. Increasing the cellular concentration of either of these decoding molecules for termination or selenocysteine incorporation showed that they were able to compete for UGA by a kinetic competition that is dynamic and dependent on the Escherichia coli growth rate. The tRNA(Sec)-mediated decoding can compete more effectively for the UGA recoding site at lower growth rates, consistent with anaerobic induction of fdhF expression.
Mansell
J B
JB
Department of Biochemistry and Centre for Gene Research, University of Otago, PO Box 56, Dunedin, New Zealand.
Guévremont
D
D
Poole
E S
ES
Tate
W P
WP
D001483:Base Sequence; D004926:Escherichia coli / Q000201:enzymology*; D005560:Formate Dehydrogenases / Q000235:genetics*; D006864:Hydrogenase / Q000235:genetics*; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes / Q000235:genetics*; D009690:Nucleic Acid Conformation; D010454:Peptide Termination Factors / Q000378:metabolism*; D014176:Protein Biosynthesis; D012345:RNA, Transfer, Amino Acid-Specific / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D009097:Multienzyme Complexes; D010454:Peptide Termination Factors; D012345:RNA, Transfer, Amino Acid-Specific; C041460:peptide chain termination release factor 2; C036928:tRNA, selenocysteine-; D006864:Hydrogenase; D005560:Formate Dehydrogenases; C054017:formate hydrogenlyase