Multiple replication factors augment DNA synthesis by the two eukaryotic DNA polymerases, alpha and delta

Tsurimoto, T., Stillman, B. (December 1989) Multiple replication factors augment DNA synthesis by the two eukaryotic DNA polymerases, alpha and delta. Embo J, 8 (12). pp. 3883-9. ISSN 0261-4189



DNA synthesis by two eukaryotic DNA polymerases, alpha and delta, was studied using a single-strand M13 DNA template primed at a unique site. In the presence of low amounts of either DNA polymerase alpha or delta, DNA synthesis was limited and short DNA strands of approximately 100 bases were produced. Addition of replication factors RF-A, PCNA and RF-C, which were previously shown to be required for SV40 DNA replication in vitro, differentially stimulated the activity of both DNA polymerases. RF-A and RF-C independently stimulated DNA polymerase alpha activity 4- to 6-fold, yielding relatively short DNA strands (less than 1 kb) and PCNA had no effect. In contrast, polymerase delta activity was stimulated co-operatively by PCNA, RF-A and RF-C approximately 25- to 30-fold, yielding relatively long DNA strands (up to 4 kb). Neither RF-C nor RF-A appear to correspond to known polymerase stimulatory factors. RF-A was previously shown to be required for initiation of DNA replication at the SV40 origin. Results presented here suggest that it also functions during elongation. The differential effects of these three replication factors on DNA polymerases alpha and delta is consistent with the model that the polymerases function at the replication fork on the lagging and leading strand templates respectively. We further suggest that co-ordinated synthesis of these strands requires dynamic protein-protein interactions between these replication factors and the two DNA polymerases.

Item Type: Paper
Uncontrolled Keywords: Bacterial Proteins pharmacology DNA Helicases DNA Polymerase I physiology DNA Polymerase II physiology DNA Replication DNA Bacterial biosynthesis DNA Viral biosynthesis DNA-Binding Proteins metabolism Models Genetic Nuclear Proteins metabolism Poly dA-dT Proliferating Cell Nuclear Antigen Proteins Simian virus 40 genetics Trans-Activators
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > enzymes > DNA polymerase
CSHL Authors:
Communities: CSHL labs > Stillman lab
Highlight: Stillman, Bruce W.
Depositing User: CSHL Librarian
Date: 1 December 1989
Date Deposited: 28 Feb 2012 19:38
Last Modified: 20 Jun 2017 20:17
PMCID: PMC402077

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