Comparison among DNA polymerases 1, 2 and 3 from maize embryo axes. A DNA primase activity copurifies with DNA polymerase 2.
Three DNA polymerase activities, named 1, 2 and 3 were purified from maize embryo axes and were compared in terms of ion requirements, optimal pH, temperature and KCl for activity, response to specific inhibitors and use of templates. All three enzymes require a divalent cation for activity, but main differences were observed in sensitivity to inhibitors and template usage: while DNA polymerases 1 and 2 were inhibited by N-ethyl maleimide and aphidicolin, inhibitors of replicative-type enzymes, DNA polymerase 3 was only marginally or not affected at all. In contrast, DNA polymerase 3 was highly inhibited by very low concentrations of ddTTP, an inhibitor of repair-type enzymes, and a 100-fold higher concentration of the drug was needed to inhibit DNA polymerases 1 and 2. Additionally, DNA polymerases 1 and 2 used equally or more efficiently the synthetic template polydA-oligodT, as compared to activated DNA, while polymerase 3 used it very poorly. Whereas DNA polymerases 1 and 2 shared properties of replicative-type enzymes, DNA polymerase 3 could be a repair-type enzyme. Moreover, a DNA primase activity copurified with the 8000-fold purified DNA polymerase 2, strengthening the suggestion that polymerase 2 is a replicative enzyme, of the alpha-type. This DNA primase activity was also partially characterized. The results are discussed in terms of relevant data about other plant DNA polymerases and primases reported in the literature.
García
E
E
Departamento de Bioquimica, Facultad de Quimica, Universidad y Copilco, Mexico D.F.
Orjuela
D
D
Camacho
Y
Y
Zúñiga
J J
JJ
Plasencia
J
J
Vázquez-Ramos
J M
JM
D004257:DNA Polymerase II / Q000302:isolation & purification* / Q000378:metabolism; D019915:DNA Primase; D004259:DNA-Directed DNA Polymerase / Q000302:isolation & purification* / Q000378:metabolism; D018525:Germination; D008274:Magnesium / Q000378:metabolism; D019384:Nucleic Acid Synthesis Inhibitors; D009994:Osmolar Concentration; D012316:RNA Nucleotidyltransferases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D013696:Temperature; D013698:Templates, Genetic; D003313:Zea mays / Q000196:embryology* / Q000201:enzymology* / Q000378:metabolism
D019384:Nucleic Acid Synthesis Inhibitors; D019915:DNA Primase; D012316:RNA Nucleotidyltransferases; D004257:DNA Polymerase II; D004259:DNA-Directed DNA Polymerase; D008274:Magnesium