Abstract
The interaction of DNA polymerase δ purified from eggs of the teleost fish Misgurnus fossilis (loach) with DNA duplexes with single-strand gaps of 1-13 nucleotides was studied. In the absence of template-restricting DNA, the enzyme elongated primers on single-stranded DNA templates in a distributive manner. However, in the presence of the proximal 5"-terminus restricting the template, the enzyme activity significantly increased. In this case, the enzyme was capable of processive synthesis by filling gaps of 5-9 nucleotides in DNA duplexes. These data indicate that DNA polymerase δ can interact with both the 3"- and 5"-termini located upstream and downstream from the gap. Analysis of the complexes formed by DNA polymerase δ and different DNA substrates by electrophoretic mobility shift assay confirmed the assumption that this enzyme can interact with the proximal 5"-terminus restricting the gap. DNA polymerase δ displayed much higher affinity in duplexes with gaps of approximately 10 nucleotides compared to the standard template–primer complexes. Maximal affinity was observed in experiments with DNA substrates containing unpaired 3"-tails in primers. The results of this study suggest that DNA polymerase δ exerts high activity in the cell nuclei during repair of DNA intermediates with single strand gaps and unpaired 3"-termini.
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Sharova, N.P., Abramova, E.B., Dmitrieva, S.B. et al. Interaction of Loach DNA Polymerase δ with DNA Duplexes with Single-Strand Gaps. Biochemistry (Moscow) 66, 402–409 (2001). https://doi.org/10.1023/A:1010297312279
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DOI: https://doi.org/10.1023/A:1010297312279