title: Stretching DNA to quantify nonspecific protein binding.
abstract: Nonspecific binding of regulatory proteins to DNA can be an important mechanism for target search and storage. This seems to be the case for the lambda repressor protein (CI), which maintains lysogeny after infection of E. coli. CI binds specifically at two distant regions along the viral genome and induces the formation of a repressive DNA loop. However, single-molecule imaging as well as thermodynamic and kinetic measurements of CI-mediated looping show that CI also binds to DNA nonspecifically and that this mode of binding may play an important role in maintaining lysogeny. This paper presents a robust phenomenological approach using a recently developed method based on the partition function, which allows calculation of the number of proteins bound nonspecific to DNA from measurements of the DNA extension as a function of applied force. This approach was used to analyze several cycles of extension and relaxation of λ DNA performed at several CI concentrations to measure the dissociation constant for nonspecific binding of CI (~100 nM), and to obtain a measurement of the induced DNA compaction (~10%) by CI.
authors.affiliation: Department of Physics, Emory University, Atlanta, Georgia 30322, USA.
authors.forename: Sachin
authors.initials: S
authors.lastname: Goyal
authors.forename: Chandler
authors.initials: C
authors.lastname: Fountain
authors.forename: David
authors.initials: D
authors.lastname: Dunlap
authors.forename: Fereydoon
authors.initials: F
authors.lastname: Family
authors.forename: Laura
authors.initials: L
authors.lastname: Finzi
mesh_terms: D003198:Computer Simulation; D004247:DNA / Q000737:chemistry* / Q000648:ultrastructure; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000648:ultrastructure; D008846:Micromanipulation / Q000379:methods*; D008956:Models, Chemical; D015336:Molecular Probe Techniques; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods*
chemical_list: D004268:DNA-Binding Proteins; D004247:DNA