DNA polymerase III from Saccharomyces cerevisiae. II. Inhibitor studies and comparison with DNA polymerase I and II

P. M.J. Burgers, G. A. Bauer

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Abstract

The newly identified yeast DNA polymerase III was compared to DNA polymerases I and II and the mitochondrial DNA polymerase. Inhibition by aphidicolin (I50) of DNA polymerases I, II, and III was 4, 6, and 0.6 μg/ml, respectively. The mitochondrial enzyme was insensitive to the drug. N2-(p-n-butylphenyl)-2'-deoxyguanosine 5'-triphosphate strongly inhibited DNA polymerase I (I50 = 0.3 μM), whereas DNA polymerase III was less sensitive (I50 = 80 μM). Conditions that allowed proteolysis to proceed during the preparation of extracts converted DNA polymerase II from a sensitive form (I50 = 2.4 μM) to a resistant form (I50 = 2 mM). The mitochondrial DNA polymerase is insensitive (I50 > 5 mM). With most other inhibitors tested (N-ethylmaleimide, heparin, salt) only small differences were observed between the three nuclear DNA polymerases. Polyclonal antibodies to DNA polymerase III did not inhibit DNA polymerases I and II, nor were those polymerases recognized by Western blotting. Monoclonal antibodies to DNA polymerase I did not cross-react with DNA polymerases II and III. The results show that DNA polymerase III is distinct from DNA polymerases I and II.

Original languageEnglish
Pages (from-to)925-930
Number of pages6
JournalJournal of Biological Chemistry
Volume263
Issue number2
StatePublished - 1988

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