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Exonucleases are enzymes (found as individual enzymes, or as parts of larger enzyme complexes) that cleave nucleotides one at a time from an end of a polynucleotide chain. These enzymes hydrolyze phosphodiester bonds from either the 3' or 5' terminus of a polynucleotide molecule.


DNA polymerase I contains a 5' exonuclease that clips off the RNA primer contained immediately upstream from the site of DNA synthesis in a 5' --> 3' manner. Pol I then synthesizes DNA nucleotides in place of the RNA primer it had just removed. DNA polymerase I also has 3' to 5' exonuclease activity which is used in editing and proofreading DNA for errors.

In both archaebacteria and eukaryotes, one of the main routes of RNA degradation is performed by the multi-protein exosome complex, which consists largely of 3' to 5' exoribonucleases.

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