PMID: 8602353Feb 1, 1996Paper

Oligonucleotide facilitators may inhibit or activate a hammerhead ribozyme

Nucleic Acids Research
E Jankowsky, B Schwenzer

Abstract

Facilitators are oligonucleotides capable of affecting hammerhead ribozyme activity by interacting with the substrate at the termini of the ribozyme. Facilitator effects were determined in vitro using a system consisting of a ribozyme with 7 nucleotides in every stem sequence and two substrates with inverted facilitator binding sequences. The effects of 9mer and 12mer RNA as well as DNA facilitators which bind either adjacent to the 3'- or 5'-end of the ribozyme were investigated. A kinetic model was developed which allows determination of the apparent dissociation constant of the ribozyme-substrate complex from single turnover reactions. We observed a decreased dissociation constant of the ribozyme-substrate complex due to facilitator addition corresponding to an additional stabilization energy of delta delta G=-1.7 kcal/mol with 3'-end facilitators. The cleavage rate constant was increased by 3'-end facilitators and decreased by 5'-end facilitators. Values for Km were slightly lowered by all facilitators and kcat was increased by 3'-end facilitators and decreased by 5'-end facilitators in our system. Generally the facilitator effects increased with the length of the facilitators and RNA provided greater effects than DNA of th...Continue Reading

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Citations

Sep 1, 2009·Cellular and Molecular Life Sciences : CMLS·Cedric ReymondJean-Pierre Perreault
Jan 7, 1998·Bioorganic & Medicinal Chemistry·F BurlinaJ L Fourrey
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Jul 19, 2008·IUBMB Life·Anastassios VourekasDenis Drainas
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Nov 24, 1998·Antisense & Nucleic Acid Drug Development·H A WielandB Krist
Dec 8, 2005·Proceedings·C Casey Cunningham

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