Characterization of the mitochondrial DNA polymerase from Saccharomyces cerevisiae

• Autorzy: Sen S. , Mukhopadhyay S. , Wetzel J. , Biswas T.K.
• Języki publikacji: EN

Abstrakty

EN

The mitochondrial DNA (mtDNA) polymerase was isolated from a protease-deficient yeast strain (PY2), and purified about 3000 fold by a column chromatography on phosphocellulose, heparin-agarose, and single-stranded DNA cellulose. The purified polymerase was characterized with respect to optimal nucleotide concentration, template-primer specificity and sensitivity to some inhibitors. These results were compared with the nuclear DNA polymerase I activity. Both polymerases showed similar requirement of deoxynucleotide concentrations (Km < 1 uM), and highest activity with poly(dA-dT) template. However, the mtDNA polymerase was more sensitive to ddTTP, EtBr and Mn2+ inhibition in comparison to the nuclear DNA polymerase I. The mtDNA polymerase did not need ATP as an energy source for in vitro DNA synthesis. This mtDNA polymerase preparation also showed 3' -> 5' exonudease activity.

Słowa kluczowe

EN

DNA polymerase; mitochondrion; Saccharomyces cerevisiae

• Wydawca: -
• Czasopismo: Acta Biochimica Polonica
• Rocznik: 1994
• Tom: 41
• Numer: 1
• Opis fizyczny: p.79-86,fig.

Twórcy

autor

Sen S.

• Wayne State University School of Medicine, Detroit, Ml 48201, USA

autor

Mukhopadhyay S.

autor

Wetzel J.

autor

Biswas T.K.

Bibliografia

• 1. Tzagoloff, A. & Myers, A.M. (1986) Genetics of mitochondrial biogenesis. Annu. Rev. Biochem. 55,249-285.
• 2. Costanzo, M.C. & Fox, T.D. (1990) Control of mitochondrial gene expression in Saccharomyces cerevisiae. Atmu. Rev. Genet. 24,91-113.
• 3. Bogenhagen, D. & Clayton, D.A. (1977) Mouse cell mitochondrial DNA molecules are selected randomly for replication throughout the cell cycle. Cell 11,719-727.
• 4. Newlon, C.S. & Fangman, W.L. (1975) Mitochondrial DNA synthesis in cell cycle mutants of Saccharomyces cerevisiae. Cell 7, 423-428.
• 5. Clayton, D.A. (1982) Replication of animal mitochondrial DNA. Cell 28,693-705.
• 6. Fangman, W.L., Henly, G., Churchill, G. & Brewer, B.J. (1989) Stable maintenance of a 35 base-pair yeast mitochondrial genome. Mol. Cell Biol. 9,1917-1921.
• 7. Wintersberger, U. Sc Blutsch, H. (1976) DNA-dependent DNA polymerase from yeast mitochondria. Eur. J. Biochem. 68,199-207.
• 8. Burgers, P.M.J. & Bauer, G.A. (1988) DNA polymerase III from Saccharomyces cerevisiae: inhibitor studies and comparison with DNA polymerases I and II. /. Biol. Chem. 263,925-930.
• 9. Biswas, T.K., Ticho, B. & Getz, G.S. (1987) In vitro characterization of the yeast mitochondrial promoter using single-base substitution mutants. /. Biol. Chem. 262,13690-13696.
• 10. Fry, M. & Loeb, L.A. (1986) Animal Cell DNA polymerases; pp. 91-101, CRC Press, Boca Raton, Florida.
• 11. Wernette, C. M. & Kaguni, L.S. (1986) A mitochondrial DNA polymerase from embryos of Dmsophila melanogaster: purification, subunit structure, and partial characterization /. Biol. Chem. 261,14764-14770.
• 12. Yarranton, G.T., Das, R. H. & Gefter, M L. (1979) Enzyme-catalyzed DNA unwinding: a DNA-dependent ATPase from Escherichia coli. j. Biol. Chem. 254,11997-12001.
• 13. Ohtani,T.,Slubata,T., lwabuchi, M., Watabc, H., lino, T. & Ando, T. (1982) ATP-dependent unwinding of double helix in closed circular DNA by RecA protein of Escherichia coli. Nature (London) 299,86-89.
• 14. Scott, J.F., Eisenberg, S., Bertsch, L.L. & Komberg, A. (1977) A mechanism of duplex DNA replication revealed by enzymatic studies of phage 0X174: Catalytic strand separation in advance of replication. Proc. Natl. Acad. Sci. U.S.A. 74,193-197.
• 15. Eichler, D.C. & Lehman, I.R. (1977) On the role of ATP in phosphodiester bond hydrolysis catalyzed by the recBC deoxyribonuclease of Escherichia coli. J. Biol. Chem. 252,499-503.
• 16. Putrament, A., Baranowska, H., Ejchart, A. & Prazmo, W. (1975) Manganese mutagenesis in yeast. Mol. Gen. Genet. 140,339-347.
• 17. Slonimski, P.P., Perodin, G. & Croft, J.H. (1968) Ethidium bromide induced mutation of yeast mitochondria: complete transformation of cells into respiratory deficient non-chromosomal "petites". Biochem. BUyhys. Res. Ccnnmun. 30, 232-239.
• 18. Waquar, M.A., Evans, M.J. & Huberman, J.A. (1978) Effect of 2'3'-dideoxythymidine-5,-tri- phosphatc on HeLa cell in vitro DNA synthesis: evidence that DNA polymerase a is the onlypolymerase required for cellular DNA replication. Nucl. Acids Res. 5,1933-1946.
• 19. Nelson, E.M., Stowers, D.J., Bayne, M L. & Benbow, R.M. (1983) Classification of DNA polymerase activities from ovaries of the frog, Xenoptts laevis. Dev. Biol. 96,11-18.
• 20. McLennan, A.G. (1980) Effect of aphidicolin and 2'3'-dideoxythymidine on mitochondrial DNA replication. Biochem. Biophys. Res. Cormrtun. 94, 116-122.
• 21. Insdorf, N.F. & Bogenhagen, D.F. (1989) DNA polymerase y from Xenopus laevis: a 3' -> 5' exonuclease is tightly associated with the DNA polymerase activity. J. Biol. Chem. 264, 21498-21503.
• 22. Kunkel, T.A. & Soni, A. (1988) Exonucleolytic proofreading enhances the fidelity of DNA synthesis by chick embryo DNA polymerase y. J. Biol Oiem. 263,4450-4459.
• 23. Kunkel, T.A. & Mosbaugh, D.W. (1989) Exonucleolytic proof-reading by a mammalian DNA polymerase y. Biochemistry 28,988-995.
• 24. Ollis, D.L., Brick, P., Hamlin, R., Xuong, N.G. Steitz, T.A. (1983) Structure of large fragment of Escherichia coli DNA polymerase I complexed with dTMP. Nature (London) 313,762-766.
• 25. Spicer, E.K., Rush, J., Fung, C, Reha-Krantz, L.J., Karam, J.D. & Königsberg, W.H. (1988) Primary structure of T4 DNA polymerase: evolutionary relatedness to eukaryotic and other prokaryotic DNA polymerases. /. Biol. Chein. 263,7478-7486.