title: Dynamics and consequences of DNA looping by the FokI restriction endonuclease.
abstract: Genetic events often require proteins to be activated by interacting with two DNA sites, trapping the intervening DNA in a loop. While much is known about looping equilibria, only a few studies have examined DNA-looping dynamics experimentally. The restriction enzymes that cut DNA after interacting with two recognition sites, such as FokI, can be used to exemplify looping reactions. The reaction pathway for FokI on a supercoiled DNA with two sites was dissected by fast kinetics to reveal, in turn: the initial binding of a protein monomer to each site; the protein-protein association to form the dimer, trapping the loop; the subsequent phosphodiester hydrolysis step. The DNA motion that juxtaposes the sites ought on the basis of Brownian dynamics to take approximately 2 ms, but loop capture by FokI took 230 ms. Hence, DNA looping by FokI is rate limited by protein association rather than DNA dynamics. The FokI endonuclease also illustrated activation by looping: it cut looped DNA 400 times faster than unlooped DNA.
authors.affiliation: The DNA-Protein Interactions Unit, Department of Biochemistry, School of Medical Sciences, University of Bristol, University Walk, Bristol BS8 1TD, UK.
authors.forename: Lucy E
authors.initials: LE
authors.lastname: Catto
authors.forename: Stuart R W
authors.initials: SR
authors.lastname: Bellamy
authors.forename: Susan E
authors.initials: SE
authors.lastname: Retter
authors.forename: Stephen E
authors.initials: SE
authors.lastname: Halford
mesh_terms: D001665:Binding Sites; D004278:DNA, Superhelical / Q000737:chemistry* / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism*; D019281:Dimerization; D007700:Kinetics; D009690:Nucleic Acid Conformation; D011485:Protein Binding
chemical_list: D004278:DNA, Superhelical; C035042:endodeoxyribonuclease FokI; D015252:Deoxyribonucleases, Type II Site-Specific