title: Two subunits of EcoRII restriction endonuclease interact with two DNA recognition sites.
abstract: The cleavage of a 14 base pair DNA duplex containing one EcoRII recognition site by EcoRII restriction endonuclease (R.EcoRII) was studied in single turnover experiments with varying enzyme concentrations in the micromolar range. The reaction rate increased with enzyme concentration until a ratio of one dimeric R.EcoRII enzyme to two double stranded oligonucleotide molecules. Excess of R.EcoRII lead to inhibition of cleavage. Maximum cleavage was also found with pBR322 DNA containing six EcoRII recognition sites at a ratio of one dimeric enzyme to two EcoRII recognition sites of the plasmid DNA. At higher ratios inhibition was observed. These observations indicate that the active enzyme complex is formed when two subunits of the enzyme interact with two R.EcoRII recognition sites.
authors.affiliation: A.N. Belozersky Institute of Physico-Chemical Biology, Moscow State University, Russia.
authors.forename: O V
authors.initials: OV
authors.lastname: Petrauskene
authors.forename: E A
authors.initials: EA
authors.lastname: Karpova
authors.forename: E S
authors.initials: ES
authors.lastname: Gromova
authors.forename: W
authors.initials: W
authors.lastname: Guschlbauer
mesh_terms: D001482:Base Composition; D001483:Base Sequence; D001665:Binding Sites; D004247:DNA / Q000378:metabolism*; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism*; D007700:Kinetics; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis / Q000378:metabolism*; D013379:Substrate Specificity
chemical_list: D046911:Macromolecular Substances; D009838:Oligodeoxyribonucleotides; D004247:DNA; C046545:CCWGG-specific type II deoxyribonucleases; D015252:Deoxyribonucleases, Type II Site-Specific