Functional interactions of mitochondrial DNA polymerase and single-stranded DNA-binding protein. Template-primer DNA binding and initiation and elongation of DNA strand synthesis
- PMID: 10329675
- DOI: 10.1074/jbc.274.21.14779
Functional interactions of mitochondrial DNA polymerase and single-stranded DNA-binding protein. Template-primer DNA binding and initiation and elongation of DNA strand synthesis
Abstract
Functional interactions between mitochondrial DNA polymerase (pol gamma) and mitochondrial single-stranded DNA-binding protein (mtSSB) from Drosophila embryos have been evaluated with regard to the overall activity of pol gamma and in partial reactions involving template-primer binding and initiation and idling in DNA strand synthesis. Both the 5' --> 3' DNA polymerase and 3' --> 5' exonuclease in pol gamma are stimulated 15-20-fold on oligonucleotide-primed single-stranded DNA by native and recombinant forms of mtSSB. That the extent of stimulation is similar for both enzyme activities over a broad range of KCl concentrations suggests their functional coordination and a similar mechanism of stimulation by mtSSB. At the same time, the high mispair specificity of pol gamma in exonucleolytic hydrolysis is maintained, indicating that enhancement of pol gamma catalytic efficiency is likely not accompanied by increased nucleotide turnover. DNase I footprinting of pol gamma.DNA complexes and initial rate measurements show that mtSSB enhances primer recognition and binding and stimulates 30-fold the rate of initiation of DNA strands. Dissociation studies show that productive complexes of the native pol gamma heterodimer with template-primer DNA are formed and remain stable in the absence of replication accessory proteins.
Similar articles
-
Effects of Xenopus laevis mitochondrial single-stranded DNA-binding protein on primer-template binding and 3'-->5' exonuclease activity of DNA polymerase gamma.J Biol Chem. 1996 Aug 2;271(31):18939-46. doi: 10.1074/jbc.271.31.18939. J Biol Chem. 1996. PMID: 8702557
-
Physiological and biochemical defects in functional interactions of mitochondrial DNA polymerase and DNA-binding mutants of single-stranded DNA-binding protein.J Biol Chem. 2004 Apr 23;279(17):17047-53. doi: 10.1074/jbc.M400283200. Epub 2004 Jan 30. J Biol Chem. 2004. PMID: 14754882
-
Mitochondrial Single-stranded DNA-binding Proteins Stimulate the Activity of DNA Polymerase γ by Organization of the Template DNA.J Biol Chem. 2015 Nov 27;290(48):28697-707. doi: 10.1074/jbc.M115.673707. Epub 2015 Oct 7. J Biol Chem. 2015. PMID: 26446790 Free PMC article.
-
DNA polymerase gamma, the mitochondrial replicase.Annu Rev Biochem. 2004;73:293-320. doi: 10.1146/annurev.biochem.72.121801.161455. Annu Rev Biochem. 2004. PMID: 15189144 Review.
-
Replicating animal mitochondrial DNA.Genet Mol Biol. 2013 Sep;36(3):308-15. doi: 10.1590/S1415-47572013000300002. Epub 2013 Aug 30. Genet Mol Biol. 2013. PMID: 24130435 Free PMC article. Review.
Cited by
-
Structural and Molecular Basis for Mitochondrial DNA Replication and Transcription in Health and Antiviral Drug Toxicity.Molecules. 2023 Feb 14;28(4):1796. doi: 10.3390/molecules28041796. Molecules. 2023. PMID: 36838782 Free PMC article. Review.
-
Unravelling How Single-Stranded DNA Binding Protein Coordinates DNA Metabolism Using Single-Molecule Approaches.Int J Mol Sci. 2023 Feb 1;24(3):2806. doi: 10.3390/ijms24032806. Int J Mol Sci. 2023. PMID: 36769124 Free PMC article. Review.
-
Mechanism of strand displacement DNA synthesis by the coordinated activities of human mitochondrial DNA polymerase and SSB.Nucleic Acids Res. 2023 Feb 28;51(4):1750-1765. doi: 10.1093/nar/gkad037. Nucleic Acids Res. 2023. PMID: 36744436 Free PMC article.
-
Mitochondrial DNA maintenance in Drosophila melanogaster.Biosci Rep. 2022 Nov 30;42(11):BSR20211693. doi: 10.1042/BSR20211693. Biosci Rep. 2022. PMID: 36254835 Free PMC article. Review.
-
Polymerase ζ Is Involved in Mitochondrial DNA Maintenance Processes in Concert with APE1 Activity.Genes (Basel). 2022 May 13;13(5):879. doi: 10.3390/genes13050879. Genes (Basel). 2022. PMID: 35627264 Free PMC article.
Publication types
MeSH terms
Substances
Grants and funding
LinkOut - more resources
Full Text Sources
Molecular Biology Databases