title: How the EcoRI endonuclease recognizes and cleaves DNA.
abstract: One popular recombinant DNA tool is the EcoRI endonuclease, which cleaves DNA at GAATTC sites and serves as a paradigm for sequence specific DNA-enzyme interactions. The recently revised X-ray crystal structure of an EcoRI-DNA complex reveals EcoRI employs novel DNA recognition motifs, a four alpha-helix bundle and two extended chains, which project into the major groove to contact substrate purines and pyrimidines. Interestingly, pyrimidine contacts had been predicted based on genetic and biochemical studies. Current work focuses on the EcoRI active site structure, enzyme and substrate conformational changes during catalysis, and host-restriction system interactions.
authors.affiliation: Rockefeller University, New York, NY 10021.
authors.forename: J
authors.initials: J
authors.lastname: Heitman
mesh_terms: D000494:Allosteric Regulation; D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D002384:Catalysis; D004269:DNA, Bacterial / Q000235:genetics / Q000378:metabolism*; D015246:Deoxyribonuclease EcoRI / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D011485:Protein Binding; D011487:Protein Conformation; D013379:Substrate Specificity; D014961:X-Ray Diffraction
chemical_list: D004269:DNA, Bacterial; D015246:Deoxyribonuclease EcoRI