title: [Selective inhibition of 3'-5'-exonuclease activity of DNA-polymerase I from Escherichia coli by a fluoride ion].
abstract: The effect of NaF on the enzymatic activities of the large fragment of E. coli DNA polymerase I (Klenow enzyme-KE) with different DNA-substrates was studied. It was shown that fluoride ion at concentrations of 5-10 mM efficiently inhibits the 3'----5' exonuclease activity of KE but does not affect the polymerase activity of the enzyme. Selective inhibition of the 3'----5' exonuclease activity of KE is Mg-dependent and is observed with double- or single-stranded DNAs. In reaction with the 14-mer oligonucleotide annealed with single-stranded phage M13 DNA the enzyme was found not only to perform the exonucleolytic hydrolysis of the primers but to catalyse also a limited elongation of some primers, adding a few nucleotide residues in the absence of exogenous dNTP. The primer elongation is inhibited by inorganic pyrophosphatase and is stimulated by micromolar concentrations of exogenous pyrophosphate thus suggesting a possible role of PPi contamination in dNTP generation via pyrophosphorolysis. Traces of precursors in DNA preparations obtained by generally employed methods may serve as another source of nucleotides for the primer elongation.
authors.forename: V S
authors.initials: VS
authors.lastname: Mikhaĭlov
authors.forename: D O
authors.initials: DO
authors.lastname: Ataeva
authors.forename: K A
authors.initials: KA
authors.lastname: Marlyev
authors.forename: A M
authors.initials: AM
authors.lastname: Atrazhev
mesh_terms: D001345:Autoradiography; D003090:Coliphages / Q000378:metabolism; D004247:DNA / Q000096:biosynthesis; D004256:DNA Polymerase I / Q000037:antagonists & inhibitors* / Q000378:metabolism; D011756:Diphosphates / Q000494:pharmacology; D004926:Escherichia coli / Q000201:enzymology*; D005092:Exonucleases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D006868:Hydrolysis; D012969:Sodium Fluoride / Q000494:pharmacology*; D013379:Substrate Specificity
chemical_list: D011756:Diphosphates; D012969:Sodium Fluoride; D004247:DNA; D004256:DNA Polymerase I; D005092:Exonucleases
vernacular_title: Selektivnoe ingibirovanie ftorid-ionom 3'-5'-ékzonukleaznoĭ aktivnosti DNK-polimerazy I Escherichia coli.