Helicases are ubiquitous enzymes involved in all aspects of DNA metabolism including replication, repair, recombination, and transcription. The mechanism of the bacteriophage T4 Dda helicase was investigated by preparing a DNA-PNA chimeric substrate. Surprisingly, Dda was able to unwind a substrate containing 12 PNA moieties in the loading strand of the enzyme. We suggest a mechanism whereby the Dda helicase contains two distinct DNA binding domains which allow an inchworm mechanism for translocation. A single step of the enzyme is sufficient to unwind the DNA-PNA chimera because several base pairs melt spontaneously due to thermal fraying. Hence, Dda helicase can unwind the substrate without actually translocating along the PNA.
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