Abstract
E. coli RecBCD is a bipolar DNA helicase consisting of two superfamily-1 motor subunits: RecB (3’ to 5’ directionality) and RecD (5’ to 3’ directionality). Although these subunits have opposite translocation polarities, they function in unison and unwind duplex DNA in the same net direction by acting on opposite ends of the nucleic acid. We have investigated previously the mechanism of initiation of DNA unwinding by the single motor RecBC helicase, which lacks the RecD motor subunit. In order to understand the relationship between single stranded (ss) DNA translocation and DNA unwinding, we compared the ssDNA translocation mechanisms of the RecB monomer and the RecBC heterodimer using stopped-flow fluorescence approaches.
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