Preferential interaction of loach DNA polymerase δ with DNA duplexes containing single-stranded gaps
| Autor: | Natalya P Sharova, Victor S. Mikhailov, S. B. Dmitrieva, E. B. Abramova, Diana D Dimitrova |
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| Rok vydání: | 2000 |
| Předmět: |
Oocyte
Saccharomyces cerevisiae Proteins DNA polymerase DNA repair DNA polymerase II Biophysics Replication Biochemistry DNA polymerase delta Substrate Specificity Minor Histocompatibility Antigens chemistry.chemical_compound Adenosine Triphosphate Structural Biology Proliferating Cell Nuclear Antigen Gapped DNA Genetics Animals Replication Protein C Molecular Biology DNA Polymerase III Sequence Deletion Homeodomain Proteins biology Hydrolysis DNA replication DNA Templates Genetic DNA polymerase δ Cell Biology Processivity Molecular biology DNA-Binding Proteins Repressor Proteins Cypriniformes Proto-Oncogene Proteins c-bcl-2 chemistry biology.protein Primer (molecular biology) DNA Probes Teleost fish Protein Binding |
| Zdroj: | FEBS Letters. 486:14-18 |
| ISSN: | 0014-5793 |
| DOI: | 10.1016/s0014-5793(00)02232-8 |
| Popis: | We studied the interaction of DNA polymerase delta (pol delta) purified from the eggs of the teleost fish Misgurnus fossilis (loach) with DNA duplexes containing single-stranded gaps of 1-13 nucleotides (nt). In the absence of processivity factors (PCNA, RF-C, and ATP), pol delta elongated primers on single-stranded DNA templates in a distributive manner. However, the enzyme was capable of processive synthesis by filling gaps of 5-9 nt in DNA duplexes. These data suggest that, upon filling a small gap, pol delta interacts with the 5'-terminus downstream of the gap as well as with the 3'-terminus of the primer. Interaction of pol delta with the proximal 5'-terminus restricting the gap was confirmed by electrophoretic mobility shift assay. Analysis of the enzyme binding to DNA duplexes containing gaps of various sizes showed a much higher affinity of pol delta for duplexes with gaps of about 10 nt than for DNA substrates with primers annealed to single-stranded templates. The most efficient pol delta binding was observed in experiments with DNA substrates containing unpaired 3'-tails in primers. The data obtained suggest that DNA molecules with small gaps and single-stranded tails may serve as substrates for direct action of pol delta in the course of DNA repair. |
| Databáze: | OpenAIRE |
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