Direct Interaction between Mammalian DNA Polymerase β and Proliferating Cell Nuclear Antigen
| Autor: | Samuel H. Wilson, Padmini S. Kedar, Anthony J. Robertson, Esther W. Hou, Soon-Jong Kim, Rajendra Prasad, Julie K. Horton |
|---|---|
| Rok vydání: | 2002 |
| Předmět: |
DNA polymerase
DNA repair Immunoprecipitation Amino Acid Motifs Molecular Sequence Data Mutant Biochemistry Cell Line Mice Structure-Activity Relationship Proliferating Cell Nuclear Antigen Two-Hybrid System Techniques hemic and lymphatic diseases Animals Amino Acid Sequence Molecular Biology DNA Polymerase beta Binding Sites biology DNA replication DNA Cell Biology Precipitin Tests Molecular biology In vitro Proliferating cell nuclear antigen Cell biology biology.protein Sequence motif |
| Zdroj: | Journal of Biological Chemistry. 277:31115-31123 |
| ISSN: | 0021-9258 |
| DOI: | 10.1074/jbc.m201497200 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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