HIGH-ANGLE AND LIGAND-INDUCED LOW-ANGLE DNA BENDS INCITED BY OCCR LIEIN THE SAME PLANE WITH OCCR BOUND TO THE INTERIOR ANGLE

The OccR protein of Agrobacterium tumefaciens activates transcription of the occQ operon in response to octopine. Octopine shortens the DNas e I footprint of OccR and relaxes an OccR-incited DNA bend. In this st udy, we used hydroxyl radical footprinting to show that OccR contacts only one face of the operator. This contact spans 50 nucleotide pairs in the absence of octopine, and 39 nucleotide pairs in the presence of octopine. Octopine enhanced protection of the central region of the f ootprint. OccR blocked intercalation by methidiumpropyl EDTA along mos t of the operator, but did not block intercalation at the bend center. OccR protected DNA against both reagents far more completely at the p romoter-distal half of the operator than at the promoter-promixal half . Experiments to alter the phasing between the operator bend and an ad jacent sequence-directed bend indicated that the high angle DNA bend i n the absence of octopine, and the low angle bend in the presence of o ctopine, lie in the same plane. Operator DNA was bent toward the helic al face that OccR protected from hydroxyl radicals, indicating that Oc cR occupies the interior angle of this bend. (C) 1995 Academic Press L imited