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The second two activities of DNA Pol I are important for replication, but DNA Polymerase III (Pol III) is the enzyme that performs the 5'-3' polymerase function. 1  Primase-The requirement for a free 3' hydroxyl group is fulfilled by the RNA primers that are synthesized at the initiation sites by these enzymes.he second two activities of DNA Pol I are important for replication, but DNA Polymerase III (Pol III) is the enzyme that performs the 5'-3' polymerase function. 1  Primase-The requirement for a free 3' hydroxyl group is fulfilled by the RNA primers that are synthesized at the initiation sites by these enzymes.
What is the function of DNA polymerase I? Proofreads DNA. (about 20 at a time before it dissociates). It is a 5' to 3' DNA polymerase but also has 5' to 3' and 3' to 5' exonuclease activity to proof read and correct mismatches. It works with DNA:DNA as well as RNA:DNA.