title: Highly selective affinity labeling of DNA polymerase alpha-primase from human placenta by reactive analogs of ATP.
abstract: Highly selective affinity labeling of a DNA-polymerase alpha-primase complex from human placenta by o-formylphenyl esters of ATP, ADP and AMP was performed in a two-step procedure in which a substrate analog attached to the active center was elongated by radioactive ATP. If the covalent attachment is performed in the presence of poly(dT) template, the ATP esters modify selectively the delta subunit of the complex. If poly(dT) is added after the covalent binding of the reagent, both delta and gamma subunits become labeled. With the o-formylphenyl ester of AMP the delta-subunit is modified. The ADP ester modifies both the delta and gamma subunit in the presence and absence of template. It is shown that formylphenyl ester of ATP is not the substrate in the reaction of elongation catalyzed by primase. The data obtained suggest the binding site of initiating substrate to be located in the region of contact of the two subunits of primase. The role of the template in the formation of the active site is discussed.
authors.affiliation: Novosibirsk Institute of Bioorganic Chemistry, Siberian Division of the Russian Academy of Sciences.
authors.forename: O D
authors.initials: OD
authors.lastname: Zakharova
authors.forename: V N
authors.initials: VN
authors.lastname: Podust
authors.forename: A A
authors.initials: AA
authors.lastname: Mustaev
authors.forename: R O
authors.initials: RO
authors.lastname: Anarbaev
authors.forename: O I
authors.initials: OI
authors.lastname: Lavrik
mesh_terms: D000255:Adenosine Triphosphate / Q000031:analogs & derivatives*; D000345:Affinity Labels; D019915:DNA Primase; D004259:DNA-Directed DNA Polymerase; D006801:Humans; D009841:Oligonucleotides / Q000096:biosynthesis; D010920:Placenta / Q000201:enzymology*; D011061:Poly A / Q000378:metabolism; D012316:RNA Nucleotidyltransferases / Q000378:metabolism
chemical_list: D000345:Affinity Labels; D009841:Oligonucleotides; D011061:Poly A; D000255:Adenosine Triphosphate; D019915:DNA Primase; D012316:RNA Nucleotidyltransferases; D004259:DNA-Directed DNA Polymerase