In vitro activity of the hairpin ribozyme derived from the negative strand of arabis mosaic virus satellite RNA

Shin Hisamatsu, Yuki Morikawa, Rie Tomita, Terumichi Tanaka, Shigenori Sonoki, Yo Kikuchi

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

The negative strand of the satellite RNA of tobacco ringspot virus [(-)sTRSV] is a self-cleaving RNA, of which self-cleaving domain is called the hairpin ribozyme. The negative strand of the satellite RNA of arabis mosaic virus [(-)sArMV] has been suggested to have a hairpin ribozyme-like secondary structure, and we have previously shown that this hairpin domain of(-)sArMV has ribozyme activity. Here we report characterization of the cleavage reaction of the (-)sArMV hairpin ribozyme. Mutagenesis analyses in a transacting system revealed, surprisingly, that the wild-type ribozyme was less active than almost all the other mutant ribozymes tested. In a cis-acting system (self-cleaving reaction), however, the reaction of the RNA containing the wild-type sequence proceeds highly efficiently. This result suggests that the inefficient cleavage of the wild-type substrate in trans-acting system may be due to low efficiency at the substrate-binding step but not at the chemical cleavage step in the reaction. We also constructed a chimeric ribozyme between the catalytic hairpin domain from (-)sArMV and the substrate-binding site from (-)sTRSV. This chimeric ribozyme had the highest activity among the traits-acting hairpin ribozymes tested.

Original languageEnglish
Pages (from-to)352-357
Number of pages6
JournalJournal of Biochemistry
Volume122
Issue number2
Publication statusPublished - 1997 Aug
Externally publishedYes

Fingerprint

Arabis
Satellite RNAs
Mosaic Viruses
Catalytic RNA
Viruses
Nepovirus
Substrates
RNA
Mutagenesis
Tobacco
Catalytic Domain
Binding Sites
In Vitro Techniques
hairpin ribozyme

Keywords

  • Arabis mosaic virus
  • Hairpin ribozyme
  • Ribozyme
  • RNA enzyme
  • Satellite RNA

ASJC Scopus subject areas

  • Biochemistry

Cite this

Hisamatsu, S., Morikawa, Y., Tomita, R., Tanaka, T., Sonoki, S., & Kikuchi, Y. (1997). In vitro activity of the hairpin ribozyme derived from the negative strand of arabis mosaic virus satellite RNA. Journal of Biochemistry, 122(2), 352-357.

In vitro activity of the hairpin ribozyme derived from the negative strand of arabis mosaic virus satellite RNA. / Hisamatsu, Shin; Morikawa, Yuki; Tomita, Rie; Tanaka, Terumichi; Sonoki, Shigenori; Kikuchi, Yo.

In: Journal of Biochemistry, Vol. 122, No. 2, 08.1997, p. 352-357.

Research output: Contribution to journalArticle

Hisamatsu, S, Morikawa, Y, Tomita, R, Tanaka, T, Sonoki, S & Kikuchi, Y 1997, 'In vitro activity of the hairpin ribozyme derived from the negative strand of arabis mosaic virus satellite RNA', Journal of Biochemistry, vol. 122, no. 2, pp. 352-357.
Hisamatsu S, Morikawa Y, Tomita R, Tanaka T, Sonoki S, Kikuchi Y. In vitro activity of the hairpin ribozyme derived from the negative strand of arabis mosaic virus satellite RNA. Journal of Biochemistry. 1997 Aug;122(2):352-357.
Hisamatsu, Shin ; Morikawa, Yuki ; Tomita, Rie ; Tanaka, Terumichi ; Sonoki, Shigenori ; Kikuchi, Yo. / In vitro activity of the hairpin ribozyme derived from the negative strand of arabis mosaic virus satellite RNA. In: Journal of Biochemistry. 1997 ; Vol. 122, No. 2. pp. 352-357.
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