Enhancement in the cleavage activity of a hammerhead ribozyme by cationic comb-type polymers and an RNA helicase in vitro. 2002

Hideo Akashi, and Hiroaki Kawasaki, and Won Jong Kim, and Toshihiro Akaike, and Kazunari Taira, and Atsushi Maruyama
Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo, Hongo, Tokyo 113-8656, Japan.

The activity of a hammerhead ribozyme (Rz) in vivo depends on several factors, such as abundance, stability, and accessibility of Rz to its target mRNA. Among these factors, accessibility is believed to be the rate-limiting factor for Rz-mediated cleavage in vivo. As Rz and its substrate RNA are negatively charged, we examined whether cellular RNA-interacting proteins or artificial polycations might improve the accessibility of Rz to its substrate RNA. Specifically, we examined the effects of two kinds of cationic comb-type copolymer, alphaPLL-g-Dex, and a cellular RNA helicase on the accessibility of Rz to a model structured RNA in vitro. The cleavage activity of Rz was slightly enhanced by alphaPLL-g-Dex, probably due to an acceleration of the association/dissociation rate. And also, the RNA helicase-bound hybrid-Rz could cleave the target substrate at a significantly higher rate due to its unwinding activity for the duplex RNA substrate. These approaches should be useful in the development of efficient gene-inactivating reagents in the post-genomic era.

UI MeSH Term Description Entries
D007700 Kinetics The rate dynamics in chemical or physical systems.
D008969 Molecular Sequence Data Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories. Sequence Data, Molecular,Molecular Sequencing Data,Data, Molecular Sequence,Data, Molecular Sequencing,Sequencing Data, Molecular
D009690 Nucleic Acid Conformation The spatial arrangement of the atoms of a nucleic acid or polynucleotide that results in its characteristic 3-dimensional shape. DNA Conformation,RNA Conformation,Conformation, DNA,Conformation, Nucleic Acid,Conformation, RNA,Conformations, DNA,Conformations, Nucleic Acid,Conformations, RNA,DNA Conformations,Nucleic Acid Conformations,RNA Conformations
D009841 Oligonucleotides Polymers made up of a few (2-20) nucleotides. In molecular genetics, they refer to a short sequence synthesized to match a region where a mutation is known to occur, and then used as a probe (OLIGONUCLEOTIDE PROBES). (Dorland, 28th ed) Oligonucleotide
D011073 Polyamines Amine compounds that consist of carbon chains or rings containing two or more primary amino groups. Polyamine
D002593 Cetrimonium Compounds Cetyltrimethylammonium compounds that have cationic detergent, antiseptic, and disinfectant activities. They are used in pharmaceuticals, foods, and cosmetics as preservatives; on skin, mucous membranes, etc., as antiseptics or cleansers, and also as emulsifiers. These compounds are toxic when used orally due to neuromuscular blockade. Cetyltrimethylammonium Compounds,Cetrimides,Compounds, Cetrimonium,Compounds, Cetyltrimethylammonium
D004305 Dose-Response Relationship, Drug The relationship between the dose of an administered drug and the response of the organism to the drug. Dose Response Relationship, Drug,Dose-Response Relationships, Drug,Drug Dose-Response Relationship,Drug Dose-Response Relationships,Relationship, Drug Dose-Response,Relationships, Drug Dose-Response
D000071228 Polyelectrolytes Naturally occurring or artificially made water-soluble POLYMERS whose repeating units are ionizable. Polyelectrolytes demonstrate attributes that are typical of salts, such as electrical conductivity, and typical of polymers, such as viscosity. Conjugated Polyelectrolyte,Polyelectrolyte,Conjugated Polyelectrolytes,Polyelectrolyte, Conjugated,Polyelectrolytes, Conjugated
D000077286 Cetrimonium Cetyltrimethylammonium compound whose salts and derivatives are used primarily as topical antiseptics. 1-Hexadecyltrimethylammonium Chloride,CTAB,CTAOH,Cetavlon,Cetrimide,Cetriminium,Cetrimonium Bromide,Cetrimonium Chloride,Cetrimonium Hydroxide,Cetrimonium Iodide,Cetrimonium Methosulfate,Cetrimonium Methyl Sulfate,Cetrimonium Monosulfate,Cetyltrimethylammonium Bromide,Cetyltrimethylammonium Chloride,HTAB Cpd,Hexadecyl Trimethyl Ammonium Bromide,Hexadecyl(trimethyl)azanium,Hexadecyltrimethylammonium Bromide,Hexadecyltrimethylammonium Octylsulfonate,Octylsulfonate, Hexadecyltrimethylammonium
D001483 Base Sequence The sequence of PURINES and PYRIMIDINES in nucleic acids and polynucleotides. It is also called nucleotide sequence. DNA Sequence,Nucleotide Sequence,RNA Sequence,DNA Sequences,Base Sequences,Nucleotide Sequences,RNA Sequences,Sequence, Base,Sequence, DNA,Sequence, Nucleotide,Sequence, RNA,Sequences, Base,Sequences, DNA,Sequences, Nucleotide,Sequences, RNA

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