Mutational analysis of the length of the J3/4 domain of Escherichia coli ribonuclease P ribozyme

Shinnosuke Haga, Terumichi Tanaka, Yo Kikuchi

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

We prepared a series of length variants of the J3/4 domain of Escherichia coli ribonuclease P (RNase P) ribozyme: the four-base long J3/4 domain (A 62G63G64A65) was replaced with GGA (denoted ΔA), GA (ΔAG), A (ΔAGG), AAGGA (ΣA), AAAGGA (ΣAA), and AAAAGGA (ΣAAA). The results indicated that truncating and inserting operations of the J3/4 domain drastically reduced ribozyme activity (WT≫ ΣAA>ΣA>ΣAAA≫ΔAG>ΔA, ΔAGG), but did not affect the cleavage site selection of a substrate by the ribozyme. The reduced ribozyme activity of each mutant was rescued to some extent by the addition of a high concentration of magnesium ions. Our data indicate that the conserved AGGA sequence was important for efficient ribozyme reactions, and suggested that the length mutations affected ribozyme activity through metal ion binding steps.

Original languageEnglish
Pages (from-to)2630-2632
Number of pages3
JournalBioscience, Biotechnology and Biochemistry
Volume68
Issue number12
DOIs
Publication statusPublished - 2004 Dec
Externally publishedYes

Fingerprint

Ribonuclease P
Catalytic RNA
ribonucleases
Escherichia coli
conserved sequences
metal ions
magnesium
ions
mutation
mutants
Ions
Site selection
Conserved Sequence
Magnesium
Metal ions
Ribonucleases
Metals
Mutation
Substrates

Keywords

  • Escherichia coli
  • J3/4 domain
  • M1 RNA
  • Ribonuclease P (RNase P)
  • Ribozyme

ASJC Scopus subject areas

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

Cite this

Mutational analysis of the length of the J3/4 domain of Escherichia coli ribonuclease P ribozyme. / Haga, Shinnosuke; Tanaka, Terumichi; Kikuchi, Yo.

In: Bioscience, Biotechnology and Biochemistry, Vol. 68, No. 12, 12.2004, p. 2630-2632.

Research output: Contribution to journalArticle

Haga, Shinnosuke ; Tanaka, Terumichi ; Kikuchi, Yo. / Mutational analysis of the length of the J3/4 domain of Escherichia coli ribonuclease P ribozyme. In: Bioscience, Biotechnology and Biochemistry. 2004 ; Vol. 68, No. 12. pp. 2630-2632.
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