A dynamic competition between release factor 2 and the tRNA(Sec) decoding UGA at the recoding site of Escherichia coli formate dehydrogenase H

EMBO J. 2001 Dec 17;20(24):7284-93. doi: 10.1093/emboj/20.24.7284.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Escherichia coli / enzymology*
  • Formate Dehydrogenases / genetics*
  • Hydrogenase / genetics*
  • Molecular Sequence Data
  • Multienzyme Complexes / genetics*
  • Nucleic Acid Conformation
  • Peptide Termination Factors / metabolism*
  • Protein Biosynthesis
  • RNA, Transfer, Amino Acid-Specific / chemistry
  • RNA, Transfer, Amino Acid-Specific / genetics
  • RNA, Transfer, Amino Acid-Specific / metabolism*

Substances

  • Multienzyme Complexes
  • Peptide Termination Factors
  • RNA, Transfer, Amino Acid-Specific
  • peptide chain termination release factor 2
  • tRNA, selenocysteine-
  • Hydrogenase
  • Formate Dehydrogenases
  • formate hydrogenlyase