Competitive regulation of modular allosteric aptazymes by a small molecule and oligonucleotide effector

Hani Najafi, Michael Famulok

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

The hairpin ribozyme can catalyze the cleavage of RNA substrates by employing its conformational flexibility. To form a catalytic complex, the two domains A and B of the hairpin-ribozyme complex must interact with one another in a folding step called docking. We have constructed hairpin ribozyme variants harboring an aptamer sequence that can be allosterically induced by flavin mononucleotide (FMN). Domains A and B are separated by distinct bridge sequences that communicate the formation of the FMN-aptamer complex to domains A and B, facilitating their docking. In the presence of a short oligonucleotide that is complementary to the aptamer, catalytic activity of the ribozyme is completely abolished, due to the formation of an extended conformer that cannot perform catalysis. However, in the presence of the small molecule effector FMN, the inhibitory effect of the oligonucleotide is competitively neutralized and the ribozyme is activated 150-fold. We thus have established a new principle for the regulation of ribozyme catalysis in which two regulatory factors (an oligonucleotide and a small molecule) that switch the ribozyme's activity in opposite directions compete for the same binding site in the aptamer domain. Published by Cold Spring Harbor Laboratory Press.

Original languageEnglish
Pages (from-to)1514-1520
Number of pages7
JournalRNA
Volume11
Issue number10
DOIs
Publication statusPublished - Oct 2005
Externally publishedYes

Fingerprint

Allosteric Regulation
Catalytic RNA
Flavin Mononucleotide
Oligonucleotides
Catalysis
RNA Cleavage
Binding Sites
hairpin ribozyme

Keywords

  • Allosteric ribozyme
  • Aptazyme
  • Competitive regulation
  • Hairpin ribozyme
  • Ribozyme engineering

ASJC Scopus subject areas

  • Genetics
  • Molecular Biology

Cite this

Competitive regulation of modular allosteric aptazymes by a small molecule and oligonucleotide effector. / Najafi, Hani; Famulok, Michael.

In: RNA, Vol. 11, No. 10, 10.2005, p. 1514-1520.

Research output: Contribution to journalArticle

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