Diffusion-driven mechanisms of protein translocation on nucleic acids. 2. The Escherichia coli repressor--operator interaction: equilibrium measurements.

ABSTRACT: In this paper the equilibrium binding of lac re- pressor to operator sites has been studied as a function of monovalent salt concentration, of length of the DNA molecule containing the operator, and (by using various natural lac “pseudo”-operators) of operator base pair sequence. The nitrocellulose filter assay has been used to obtain values of repressor-operator association constants (KRO), both directly and as ratios of association to dissociation rate constants (k,/kd). Measurements of KRo have been made in the absence of MgZf or other divalent ions, allowing a direct estimate [Record, M. T., Jr., Lohman, T. M., & deHaseth, P. L. (1976) J. Mol. Biol. 107, 1451 of the contribution of electrostatic (charge-charge) interactions to the stability of the RO com- plexes. Using lac operator containing DNA restriction frag- ments of known size, we have shown the following: (i) The magnitide of the RO interaction is salt concentration de- pendent. A plot of log

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