title: Pyrovanadolysis, a pyrophosphorolysis-like reaction mediated by pyrovanadate, Mn2+, and DNA polymerase of bacteriophage T7.
abstract: DNA polymerases catalyze the 3'-5'-pyrophosphorolysis of a DNA primer annealed to a DNA template in the presence of pyrophosphate (PP(i)). In this reversal of the polymerization reaction, deoxynucleotides in DNA are converted to deoxynucleoside 5'-triphosphates. Based on the charge, size, and geometry of the oxygen connecting the two phosphorus atoms of PP(i), a variety of compounds was examined for their ability to carry out a reaction similar to pyrophosphorolysis. We describe a manganese-mediated pyrophosphorolysis-like activity using pyrovanadate (VV) catalyzed by the DNA polymerase of bacteriophage T7. We designate this reaction pyrovanadolysis. X-ray absorption spectroscopy reveals a shorter Mn-V distance of the polymerase-VV complex than the Mn-P distance of the polymerase-PP(i) complex. This structural arrangement at the active site accounts for the enzymatic activation by Mn-VV. We propose that the Mn(2+), larger than Mg(2+), fits the polymerase active site to mediate binding of VV into the active site of the polymerase. Our results may be the first documentation that vanadium can substitute for phosphorus in biological processes.
authors.affiliation: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115.
authors.forename: Barak
authors.initials: B
authors.lastname: Akabayov
authors.affiliation: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115.
authors.forename: Arkadiusz W
authors.initials: AW
authors.lastname: Kulczyk
authors.affiliation: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115.
authors.forename: Sabine R
authors.initials: SR
authors.lastname: Akabayov
authors.affiliation: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115.
authors.forename: Christopher
authors.initials: C
authors.lastname: Theile
authors.affiliation: Department of Chemistry, Merkert Chemistry Center, Boston College, Chestnut Hill, Massachusetts 02467, and.
authors.forename: Larry W
authors.initials: LW
authors.lastname: McLaughlin
authors.affiliation: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Beauchamp
authors.affiliation: Zernike Institute for Advanced Materials Centre for Synthetic Biology, Nijenborgh 4, 9747 AG Groningen, The Netherlands.
authors.forename: Antoine M
authors.initials: AM
authors.lastname: van Oijen
authors.affiliation: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115,. Electronic address: ccr@hms.harvard.edu.
authors.forename: Charles C
authors.initials: CC
authors.lastname: Richardson
mesh_terms: D017123:Bacteriophage T7 / Q000201:enzymology*; D020134:Catalytic Domain; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry*; D011756:Diphosphates / Q000737:chemistry*; D008345:Manganese / Q000737:chemistry*; D014638:Vanadates / Q000737:chemistry
chemical_list: D011756:Diphosphates; D014638:Vanadates; D008345:Manganese; C034970:bacteriophage T7 induced DNA polymerase; D004259:DNA-Directed DNA Polymerase