Novel RNA catalysts for the Michael reaction

Gerhard Sengle, Alexander Eisenführ, Paramjit S. Arora, James S. Nowick, Michael Famulok

Research output: Contribution to journalArticlepeer-review

Abstract

Background: In vitro selected ribozymes with nucleotide synthase, peptide and carbon-carbon bond forming activity provide insight into possible scenarios on how chemical transformations may have been catalyzed before protein enzymes had evolved. Metabolic pathways based on ribozymes may have existed at an early stage of evolution. Results: We have isolated a novel ribozyme that mediates Michael-adduct formation at a Michael-acceptor substrate, similar to the rate-limiting step of the mechanistic sequence of thymidylate synthase. The kinetic characterization of this catalyst revealed a rate enhancement by a factor of ∼105. The ribozyme shows substrate specificity and can act as an intermolecular catalyst which transfers the Michael-donor substrate onto an external 20-mer RNA oligonucleotide containing the Michael-acceptor system. Conclusions: The ribozyme described here is the first example of a catalytic RNA with Michael-adduct forming activity which represents a key mechanistic step in metabolic pathways and other biochemical reactions. Therefore, previously unforeseen RNA-evolution pathways can be considered, for example the formation of dTMP from dUMP. The substrate specificity of this ribozyme may also render it useful in organic syntheses.

Original languageEnglish (US)
Pages (from-to)459-473
Number of pages15
JournalChemistry and Biology
Volume8
Issue number5
DOIs
StatePublished - May 2001

Keywords

  • In vitro selection
  • Michael-addition
  • RNA catalysis
  • RNA world
  • Ribozyme

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Pharmacology
  • Drug Discovery
  • Clinical Biochemistry

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