title: Sequence-specific cleavage of RNA by a hybrid ribonuclease H.
abstract: Site-specific cleavage of the 22-, 132- and 534-base RNAs by the DNA/protein hybrid RNase H were examined. The 22-base RNA was chemically synthesized, and 132- and 534-base RNAs were prepared by run-off transcription. The hybrid enzyme cleaves these RNAs, which contain a single target sequence, primarily at the unique phosphodiester bond within the target sequence. The hybrid enzyme performs multiple turnovers, and at a substrate/enzyme ratio of 10:1 the RNAs are almost completely cleaved by the hybrid enzyme at 37 degrees C within 1 h. We propose that hybrid RNase H molecules with various oligodeoxyribonucleotides function as RNA restriction enzymes and are useful for structural and functional studies of RNA.
authors.affiliation: Protein Engineering Research Institute, Osaka, Japan.
authors.forename: C
authors.initials: C
authors.lastname: Nakai
authors.forename: A
authors.initials: A
authors.lastname: Konishi
authors.forename: Y
authors.initials: Y
authors.lastname: Komatsu
authors.forename: H
authors.initials: H
authors.lastname: Inoue
authors.forename: E
authors.initials: E
authors.lastname: Ohtsuka
authors.forename: S
authors.initials: S
authors.lastname: Kanaya
mesh_terms: D001483:Base Sequence; D001665:Binding Sites; D004247:DNA / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology; D006868:Hydrolysis; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D016297:Mutagenesis, Site-Directed; D009693:Nucleic Acid Hybridization; D012313:RNA / Q000737:chemistry / Q000378:metabolism*; D016914:Ribonuclease H / Q000235:genetics / Q000378:metabolism*; D013379:Substrate Specificity
chemical_list: D012313:RNA; D004247:DNA; D016914:Ribonuclease H