Abstract
Three DNA polymerase activities, A, B and C, were identified in extracts of exponentially growing synchronous cultures of Chlamydomonas reinhardii, and DNA polymerases A and B were characterized in detail. Both enzymes have the same binding affinity for DEAE-cellulose at pH 7.8, but can be distinguished from each other by their behaviour on phosphocellulose and DNA-agarose. 'Activated' calf thymus DNA was used as template, and the pH, K+ and bivalent-cation optima were measured. DNA polymerase A sediments at 5.3 S in glycerol gradients, with an apparent mol.wt. of 90000-100000. Polymerase B sediments between 8S and 10S in 100mM-KCl, the predominant species having an apparent mol.wt. of 200000. In 200mM-KCl, polymerase B dissociates to a single species, which sediments at 5.8S. A 3S species was found in aged preparations of both enzymes. The activity of polymerase B from cells harvested during nuclear DNA synthesis is twice that found in Chlamydomonas at other times during the cell cycle.
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- Banks G. R., Holloman W. K., Kairis M. V., Spanos A., Yarranton G. T. A DNA polymerase from Ustilago maydis. 1. Purification and properties of the polymerase activity. Eur J Biochem. 1976 Feb 2;62(1):131–142. doi: 10.1111/j.1432-1033.1976.tb10106.x. [DOI] [PubMed] [Google Scholar]
- Byrnes J. J., Downey K. M., So A. G. Bone marrow cytoplasmic deoxyribonucleic acid polymerase. Variation of pH and ionic environment as a possible control mechanism. Biochemistry. 1973 Oct 23;12(22):4378–4384. doi: 10.1021/bi00746a013. [DOI] [PubMed] [Google Scholar]
- Chandra P., Steel L. K., Laube H., Kornhuber B. Oncornaviral-like DNA polymerase activity in a case of childhood myelofibrotic syndrome. FEBS Lett. 1975 Oct 15;58(1):71–75. doi: 10.1016/0014-5793(75)80228-6. [DOI] [PubMed] [Google Scholar]
- Chang L. M., Brown M., Bollum F. J. Induction of DNA polymerase in mouse L cells. J Mol Biol. 1973 Feb 15;74(1):1–8. doi: 10.1016/0022-2836(73)90349-5. [DOI] [PubMed] [Google Scholar]
- Chang L. M. Phylogeny of DNA polymerase-beta. Science. 1976 Mar 19;191(4232):1183–1185. doi: 10.1126/science.769158. [DOI] [PubMed] [Google Scholar]
- Craig R. K., Costello P. A., Keir H. M. Dexyribonucleic acid polymerases of BHK-21/C13cells. Relationship to the physiological state of the cells, and to synchronous indution of synthesis of deoxyribonuleic acid. Biochem J. 1975 Feb;145(2):233–240. doi: 10.1042/bj1450233. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Crerar M., Pearlman R. E. Deoxyribonucleic acid polymerase from Tetrahymena pyriformis. Purification and properties of the major activity in exponentially growing cells. J Biol Chem. 1974 May 25;249(10):3123–3131. [PubMed] [Google Scholar]
- DAVIS B. J. DISC ELECTROPHORESIS. II. METHOD AND APPLICATION TO HUMAN SERUM PROTEINS. Ann N Y Acad Sci. 1964 Dec 28;121:404–427. doi: 10.1111/j.1749-6632.1964.tb14213.x. [DOI] [PubMed] [Google Scholar]
- Gefter M. L. DNA replication. Annu Rev Biochem. 1975;44:45–78. doi: 10.1146/annurev.bi.44.070175.000401. [DOI] [PubMed] [Google Scholar]
- Gillham N. W. Genetic analysis of the chloroplast and mitochondrial genomes. Annu Rev Genet. 1974;8:347–391. doi: 10.1146/annurev.ge.08.120174.002023. [DOI] [PubMed] [Google Scholar]
- Holmes A. M., Hesslewood I. P., Johnston I. R. In vitro concersion of a calf thymus 8S DNA polymerase to a 7.3S species. Nature. 1975 May 29;255(5507):420–422. doi: 10.1038/255420a0. [DOI] [PubMed] [Google Scholar]
- Holmes A. M., Johnston I. R. DNA polymerases of eukaryotes. FEBS Lett. 1975 Dec 15;60(2):233–243. doi: 10.1016/0014-5793(75)80721-6. [DOI] [PubMed] [Google Scholar]
- Johnson U. G., Porter K. R. Fine structure of cell division in Chlamydomonas reinhardi. Basal bodies and microtubules. J Cell Biol. 1968 Aug;38(2):403–425. doi: 10.1083/jcb.38.2.403. [DOI] [PMC free article] [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- Loeb L. A. Purification and properties of deoxyribonucleic acid polymerase from nuclei of sea urchin embryos. J Biol Chem. 1969 Apr 10;244(7):1672–1681. [PubMed] [Google Scholar]
- MARTIN R. G., AMES B. N. A method for determining the sedimentation behavior of enzymes: application to protein mixtures. J Biol Chem. 1961 May;236:1372–1379. [PubMed] [Google Scholar]
- Matsukage A., Sivarajan M., Wilson S. H. Studies on DNA alpha-polymerase of mouse myeloma: partial purification and comparison of three molecular forms of the enzyme. Biochemistry. 1976 Nov 30;15(24):5305–5314. doi: 10.1021/bi00669a017. [DOI] [PubMed] [Google Scholar]
- McLennan A. G., Keir H. M. Deoxyribonucleic acid polymerases of Euglena gracilis. Primer-template utilization of and enzyme activities associated with the two deoxyribonucleic acid polymerases of high molecular weight. Biochem J. 1975 Nov;151(2):239–247. doi: 10.1042/bj1510239. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McLennan A. G., Keir H. M. Deoxyribonucleic acid polymerases of Euglena gracilis. Purification and properties of two distinct deoxyribonucleic acid polymerases of high molecular weight. Biochem J. 1975 Nov;151(2):227–238. doi: 10.1042/bj1510227. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McLennan A. G., Keir H. M. Subcellular location and growth stage dependence of the DNA polymerases of Euglena gracilis. Biochim Biophys Acta. 1975 Oct 15;407(3):253–262. doi: 10.1016/0005-2787(75)90092-1. [DOI] [PubMed] [Google Scholar]
- Ross C. A., Harris W. J. DNA polymerases from Chlamydomonas reinhardii. Further characterization, action of inhibitors and associated nuclease activities. Biochem J. 1978 Apr 1;171(1):241–249. doi: 10.1042/bj1710241. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ross C. A., Harris W. J. Deoxyribonucleic acid polymerase from Chlamydomonas reinhardii. Biochem Soc Trans. 1976;4(4):806–807. doi: 10.1042/bst0040806. [DOI] [PubMed] [Google Scholar]
- Schaller H., Nüsslein C., Bonhoeffer F. J., Kurz C., Nietzschmann I. Affinity chromatography of DNA-binding enzymes on single-stranded DNA-agarose columns. Eur J Biochem. 1972 Apr 24;26(4):474–481. doi: 10.1111/j.1432-1033.1972.tb01789.x. [DOI] [PubMed] [Google Scholar]
- Schekman R., Weiner J. H., Weiner A., Kornberg A. Ten proteins required for conversion of phiX174 single-stranded DNA to duplex form in vitro. Resolution and reconstitution. J Biol Chem. 1975 Aug 10;250(15):5859–5865. [PubMed] [Google Scholar]
- Schönherr O. T., Keir H. M. The activity of deoxyribonucleic acid polymerase in some species of algae. Biochem J. 1972 Sep;129(2):285–290. doi: 10.1042/bj1290285. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Spadari S., Weissbach A. HeLa cell R-deoxyribonucleic acid polymerases. Separation and characterization of two enzymatic activities. J Biol Chem. 1974 Sep 25;249(18):5809–5815. [PubMed] [Google Scholar]
- Stavrianopoulos J. G., Karkas J. D., Chargaff E. DNA polymerase of chicken embryo: purification and properties. Proc Natl Acad Sci U S A. 1972 Jul;69(7):1781–1785. doi: 10.1073/pnas.69.7.1781. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sueoka N., Chiang K. S., Kates J. R. Deoxyribonucleic acid replication in meiosis of Chlamydomonas reinhardi. I. Isotopic transfer experiments with a strain producing eight zoospores. J Mol Biol. 1967 Apr 14;25(1):47–66. doi: 10.1016/0022-2836(67)90278-1. [DOI] [PubMed] [Google Scholar]
- Sueoka N. MITOTIC REPLICATION OF DEOXYRIBONUCLEIC ACID IN CHLAMYDOMONAS REINHARDI. Proc Natl Acad Sci U S A. 1960 Jan;46(1):83–91. doi: 10.1073/pnas.46.1.83. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tait A., Cummings D. J. DNA-dependent DNA polymerase activities from Paramecia macronuclei. Biochim Biophys Acta. 1975 Jan 20;378(2):282–295. doi: 10.1016/0005-2787(75)90116-1. [DOI] [PubMed] [Google Scholar]
- Tait G. C., Harris W. J. A deoxyribonuclease from Chlamydomonas reinhardii. 1. Purification and properties. Eur J Biochem. 1977 May 16;75(2):357–364. doi: 10.1111/j.1432-1033.1977.tb11536.x. [DOI] [PubMed] [Google Scholar]
- Weissbach A., Baltimore D., Bollum F., Gallo R., Korn D. Nomenclature of eukaryotic DNA polymerases. Eur J Biochem. 1975 Nov 1;59(1):1–2. doi: 10.1111/j.1432-1033.1975.tb02416.x. [DOI] [PubMed] [Google Scholar]
- Weissbach A. Vertebrate DNA polymerases. Cell. 1975 Jun;5(2):101–108. doi: 10.1016/0092-8674(75)90017-3. [DOI] [PubMed] [Google Scholar]
- Wickner S., Hurwitz J. Involvement of escherichia coli dnaZ gene product in DNA elongation in vitro. Proc Natl Acad Sci U S A. 1976 Apr;73(4):1053–1057. doi: 10.1073/pnas.73.4.1053. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wintersberger E. Deoxyribonucleic acid polymerases from yeast. Further purification and characterization of DNA-dependent DNA polymerases A and B. Eur J Biochem. 1974 Dec 16;50(1):41–47. doi: 10.1111/j.1432-1033.1974.tb03871.x. [DOI] [PubMed] [Google Scholar]
- Wintersberger U. Absence of a low-molecular-weight DNA polymerase from nuclei of the yeast, Saccharomyces cerevisiae. Eur J Biochem. 1974 Dec 16;50(1):197–202. doi: 10.1111/j.1432-1033.1974.tb03888.x. [DOI] [PubMed] [Google Scholar]
- Wintersberger U., Wintersberger E. Studies on deoxyribonucleic acid polymerases from yeast. 2. Partial purification and characterization of mitochondrial DNA polymerase from wild type and respiration-deficient yeast cells. Eur J Biochem. 1970 Mar 1;13(1):20–27. doi: 10.1111/j.1432-1033.1970.tb00894.x. [DOI] [PubMed] [Google Scholar]