title: Force-driven unbinding of proteins HU and Fis from DNA quantified using a thermodynamic Maxwell relation.
abstract: Determining numbers of proteins bound to large DNAs is important for understanding their chromosomal functions. Protein numbers may be affected by physical factors such as mechanical forces generated in DNA, e.g. by transcription or replication. We performed single-DNA stretching experiments with bacterial nucleoid proteins HU and Fis, verifying that the force-extension measurements were in thermodynamic equilibrium. We, therefore, could use a thermodynamic Maxwell relation to deduce the change of protein number on a single DNA due to varied force. For the binding of both HU and Fis under conditions studied, numbers of bound proteins decreased as force was increased. Our experiments showed that most of the bound HU proteins were driven off the DNA at 6.3 pN for HU concentrations lower than 150 nM; our HU data were fit well by a statistical-mechanical model of protein-induced bending of DNA. In contrast, a significant amount of Fis proteins could not be forced off the DNA at forces up to 12 pN and Fis concentrations up to 20 nM. This thermodynamic approach may be applied to measure changes in numbers of a wide variety of molecules bound to DNA or other polymers. Force-dependent DNA binding by proteins suggests mechano-chemical mechanisms for gene regulation.
authors.affiliation: Department of Physics and Astronomy, Department of Molecular Biosciences, Northwestern University, Evanston, IL 60208, USA. botaoshaw@hotmail.com
authors.forename: Botao
authors.initials: B
authors.lastname: Xiao
authors.forename: Houyin
authors.initials: H
authors.lastname: Zhang
authors.forename: Reid C
authors.initials: RC
authors.lastname: Johnson
authors.forename: John F
authors.initials: JF
authors.lastname: Marko
mesh_terms: D004247:DNA / Q000737:chemistry* / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000378:metabolism; D035021:Factor For Inversion Stimulation Protein / Q000737:chemistry* / Q000378:metabolism; D015233:Models, Statistical; D011485:Protein Binding; D013816:Thermodynamics
chemical_list: D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D035021:Factor For Inversion Stimulation Protein; C501034:Fis protein, E coli; C116822:hns protein, E coli; D004247:DNA