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Answer by Surfbanana. Confidence votes 22. Helicase and RNA polymerase separate DNA strands by breaking the hydrogen bonds between complementary bases. Helicase parts the strands of DNA during DNA replication, and RNA polymerase parts them during transcription. The enzyme that separates DNA in called DNA helicases. There are two of them that work away from the origin of replication, creating in bubble in the DNA molecule.
The process of breaking the hydrogen bonds between the nucleotide base pairs in double-stranded DNA requires energy. To break the bonds, helicases use the energy stored in a molecule called ATP, which serves as the energy currency of cells. DNA helicases also function in other cellular processes where double-stranded DNA must be separated, including DNA repair and transcription.