title: The 3'-->5' exonuclease associated with HeLa DNA polymerase epsilon.
abstract: The 3'-->5' exonuclease activity of highly purified large form of human DNA polymerase epsilon was studied. The activity removes mononucleotides from the 3' end of an oligonucleotide with a non-processive mechanism and leaves 5'-terminal trinucleotide non-hydrolyzed. This is the case both with single-stranded oligonucleotides and with oligonucleotides annealed to complementary regions of M13DNA. However, the reaction rates with single-stranded oligonucleotides are at least ten-fold when compared to those with completely base-paired oligonucleotides. Conceivably, mismatched 3' end of an oligonucleotide annealed to M13DNA is rapidly removed and the hydrolysis is slowed down when double-stranded region is reached. The preferential removal of a non-complementary 3' end and the nonprocessive mechanism are consistent with anticipated proofreading function. In addition to the 3'-->5' exonuclease activity, an 5'-->3' exonuclease activity is often present even in relatively highly purified DNA polymerase epsilon preparates suggesting that such an activity may be an essential component for the action of this enzyme in vivo. Contrary to the 3'-->5' exonuclease activity, the 5'-->3' exonuclease is separable from the polymerase activity.
authors.affiliation: Department of Biochemistry, University of Oulu, Finland.
authors.forename: L
authors.initials: L
authors.lastname: Uitto
authors.forename: J
authors.initials: J
authors.lastname: Halleen
authors.forename: P
authors.initials: P
authors.lastname: Remes
authors.forename: T
authors.initials: T
authors.lastname: Kesti
authors.forename: J E
authors.initials: JE
authors.lastname: Syväoja
mesh_terms: D001483:Base Sequence; D004257:DNA Polymerase II; D004258:DNA Polymerase III; D004277:DNA, Single-Stranded / Q000378:metabolism; D004259:DNA-Directed DNA Polymerase / Q000302:isolation & purification / Q000378:metabolism*; D005092:Exonucleases / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D013379:Substrate Specificity
chemical_list: D004277:DNA, Single-Stranded; D004257:DNA Polymerase II; D004258:DNA Polymerase III; D004259:DNA-Directed DNA Polymerase; D005092:Exonucleases