Abstract
Human cyclin/PCNA (proliferating cell nuclear antigen) is structurally, functionally, and immunologically homologous to the calf thymus auxiliary protein for DNA polymerase delta. This auxiliary protein has been investigated as a stimulatory factor for the nuclear DNA polymerases from S. cerevisiae. Calf cyclin/PCNA enhances by more than ten-fold the ability of DNA polymerase III to replicate templates with high template/primer ratios, e.g. poly(dA).oligo(dT) (40:1). The degree of stimulation increases with the template/primer ratio. At a high template/primer ratio, i.e. low primer density, cyclin/PCNA greatly increases processive DNA synthesis by DNA polymerase III. At low template/primer ratios (e.g. poly(dA).oligo(dT) (2.5:1), where addition of cyclin/PCNA only minimally increases the processivity of DNA polymerase III, a several-fold stimulation of total DNA synthesis is still observed. This indicates that cyclin/PCNA may also increase productive binding of DNA polymerase III to the template-primer and stabilize the template-primer-polymerase complex. The activity of yeast DNA polymerases I and II is not affected by addition of cyclin/PCNA. These results strengthen the hypothesis that yeast DNA polymerase III is functionally analogous to the mammalian DNA polymerase delta.
MeSH Terms
Animals
Cattle
DNA Polymerase III/metabolism
DNA Replication
DNA-Directed DNA Polymerase/metabolism
In Vitro Techniques
Kinetics
Magnesium/physiology
Nuclear Proteins/physiology
Proliferating Cell Nuclear Antigen
Saccharomyces cerevisiae/enzymology
Substrate Specificity
Templates, Genetic
Chemicals
Nuclear Proteins
Proliferating Cell Nuclear Antigen
DNA Polymerase III
DNA-Directed DNA Polymerase
Magnesium
Authors & Affiliations
1 authors, click to expand affiliations / ORCID
Burgers P M
Department of Biological Chemistry, Washington University School of Medicine, St. Louis, MO 63110.
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