Substrate shape preference of Escherichia coli ribonuclease P ribozyme and holo enzyme using bottom-half part-shifting variants of Pre-tRNA

Terumichi Tanaka, Yasuhiro Nagai, Yo Kikuchi

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

We showed previously that the bacterial ribonuclease P (RNase P) ribozyme has substrate shape preference depending on the concentrations of catalytically important magnesium ions. The ribozyme discriminates a canonical cloverleaf precursor tRNA from a hairpin RNA with a CCA-tag sequence at low concentrations of magnesium ions. By detailed analysis of the shape preference using the bottom-half part-shifting variants of a tRNA precursor, we showed that the RNAs in a T-shape structure can be substrates for the ribozyme reactions even at low concentrations of magnesium ions, and that the RNA in a natural L-shape is the best substrate for both the ribozyme and the holo enzyme. The results also showed that the position of the bottom-half part did not affect the cleavage site selection of a substrate by the enzyme. Our results are the first kinetic evidence to show the importance of the bottom-half part of tRNA molecule, and our result also showed that the holo enzyme can discriminate substrate shape as well as the ribozyme at low concentrations of metal ions.

Original languageEnglish
Pages (from-to)1992-1994
Number of pages3
JournalBioscience, Biotechnology and Biochemistry
Volume69
Issue number10
DOIs
Publication statusPublished - 2005 Oct 23
Externally publishedYes

Fingerprint

Ribonuclease P
Catalytic RNA
ribonucleases
RNA Precursors
Escherichia coli
Enzymes
RNA
Ions
Magnesium
Substrates
enzymes
magnesium
ions
Enzyme kinetics
Site selection
Transfer RNA
metal ions
Metal ions
Metals
Ribonucleases

Keywords

  • Clover-leaf
  • Hairpin RNA
  • Ribonuclease P
  • Substrate specificity

ASJC Scopus subject areas

  • Bioengineering
  • Biotechnology
  • Biochemistry
  • Biochemistry, Genetics and Molecular Biology(all)
  • Chemistry (miscellaneous)
  • Applied Microbiology and Biotechnology
  • Food Science

Cite this

Substrate shape preference of Escherichia coli ribonuclease P ribozyme and holo enzyme using bottom-half part-shifting variants of Pre-tRNA. / Tanaka, Terumichi; Nagai, Yasuhiro; Kikuchi, Yo.

In: Bioscience, Biotechnology and Biochemistry, Vol. 69, No. 10, 23.10.2005, p. 1992-1994.

Research output: Contribution to journalArticle

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