The Mammalian DNA Polymerase δ−Proliferating Cell Nuclear Antigen−Template-Primer Complex: Molecular Characterization by Direct Binding
Citations Over TimeTop 16% of 1996 papers
Abstract
Three direct assays, polyacrylamide gel electrophoresis-band mobility shift, agarose gel electrophoresis-band mobility shift, and nitrocellulose filter binding, were established to study complexes formed among mammalian DNA polymerase delta (pol delta), proliferating cell nuclear antigen (PCNA), and synthetic oligonucleotide template-primers. In all contexts, complex formation requires simultaneous presence of pol delta, PCNA, and template-primer. Moreover, we showed in one such assay that the complex formed contains each molecular component. Nuclease protection experiments demonstrate that complex formation protects template from degradation by DNase I. The mass determined for the pol delta.PCNA.template-primer complex was about 267 kDa, consistent with the participation of one molecule of pol delta, two or three molecules of PCNA and one molecule of template-primer. PCNA alone behaved as a trimer (mass determined to be about 87 kDa). Complex could be manipulated enzymologically. Measurement of off rates demonstrates directly that PCNA stabilizes the pol delta.template-primer complex.
Related Papers
- → Clamp loading, unloading and intrinsic stability of the PCNA, β and gp45 sliding clamps of human, E. coli and T4 replicases(1996)222 cited
- → Crystal structure of the DNA polymerase processivity factor of T4 bacteriophage 1 1Edited by I. A. Wilson(2000)171 cited
- → Structure-Function Relationship of the Eukaryotic DNA Replication Factor, Proliferating Cell Nuclear Antigen(1995)170 cited
- → PCNA binding domains in all three subunits of yeast DNA polymerase δ modulate its function in DNA replication(2011)81 cited
- → Synthetic Activity of Sso DNA Polymerase Y1, an Archaeal DinB-like DNA Polymerase, Is Stimulated by Processivity Factors Proliferating Cell Nuclear Antigen and Replication Factor C(2001)56 cited