Journal of Biological Chemistry · 2002 · 126 citations · 56 references
Life SciencesBiochemistryMolecular BiologyDna RepairDna ReplicationProtein-protein InteractionsDna Polymerase FunctionNucleic Acid-protein InteractionsPolymerase Chain ReactionProtein Complex AssemblyProtein-nucleic Acid InteractionsRna Polymerase FunctionBinding AssaysCo-immunoprecipitation AnalysisCompetitive Binding AssaysDna Polymerase BetaEukaryotic Dna Polymerase FunctionImmunoprecipitation AssaysPolymerase Subunit OrganizationPolymerase-dna Binding
Proliferating cell nuclear antigen (PCNA) plays an essential role in nucleic acid metabolism as a component of the DNA replication and DNA repair machinery. As such, PCNA interacts with many proteins that have a sequence motif termed the PCNA interacting motif (PIM) and also with proteins lacking a PIM. Three regions in human and rat DNA polymerases beta (beta-pol) that resemble the consensus PIM were identified, and we show here that beta-polymerase and PCNA can form a complex both in vitro and in vivo. Immunoprecipitation experiments, yeast two-hybrid analysis, and overlay binding assays were used to examine the interaction between the two proteins. Competition experiments with synthetic PIM-containing peptides suggested the importance of a PIM in the interaction, and studies of a beta-polymerase PIM mutant, H222A/F223A, demonstrated that this alteration blocked the interaction with PCNA. The results indicate that at least one of the PIM-like sequences in beta-polymerase appears to be a functional PIM and was required in the interaction between beta-polymerase and PCNA.
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Crystal structure of the eukaryotic DNA polymerase processivity factor PCNA
Talluru S.R. Krishna, Xiang‐Peng Kong, Sonja L. Gary et al. · Cell · 1994 · 916 citations
Requirement of mammalian DNA polymerase-β in base-excision repair
Robert W. Sobol, Julie K. Horton, Ralf Kühn et al. · Nature · 1996 · 832 citations