Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1973 Sep;115(3):762–768. doi: 10.1128/jb.115.3.762-768.1973

Characteristics of Deoxyribonucleic Acid Polymerase Isolated from Spores of Rhizopus stolonifer1

Cheng-Shung Gong a, Larry D Dunkle a, James L Van Etten a
PMCID: PMC246319  PMID: 4728271

Abstract

Deoxyribonucleic acid (DNA)-dependent DNA polymerase was purified several hundredfold from germinated and ungerminated spores of the fungus Rhizopus stolonifer. The partially purified enzymes from both spore stages exhibited identical characteristics; incorporation of [3H]deoxythymidine monophosphate into DNA required Mg2+, DNA, a reducing agent, and the simultaneous presence of deoxyguanosine triphosphate, deoxycytidine triphosphate, and deoxyadenosine triphosphate. Heat-denatured and activated DNAs were better templates than were native DNAs. The buoyant density of the radioactive product of the reaction was similar to that of the template DNA. The enzyme is probably composed of a single polypeptide chain with an S value of 5.12 and an estimated molecular weight of 70,000 to 75,000. During the early stages of purification, the enzyme fraction from ungerminated spores required exogenous DNA for maximum activity, whereas the corresponding enzyme fraction from germinated spores did not require added DNA. Apparently DNA polymerase from germinated spores was more tightly bound to endogenous DNA than was the enzyme from ungerminated spores.

Full text

PDF
762

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. APOSHIAN H. V., KORNBERG A. Enzymatic synthesis of deoxyribonucleic acid. IX. The polymerase formed after T2 bacteriophage infection of Escherichia coli: a new enzyme. J Biol Chem. 1962 Feb;237:519–525. [PubMed] [Google Scholar]
  2. Brakke M. K., Van Pelt N. Linear-log sucrose gradients for estimating sedimentation coefficients of plant viruses and nucleic acids. Anal Biochem. 1970 Nov;38(1):56–64. doi: 10.1016/0003-2697(70)90155-7. [DOI] [PubMed] [Google Scholar]
  3. Brambl R. M., Van Etten J. L. Protein synthesis during fungal spore germination. V. Evidence that the ungerminated conidiospores of Botryodiplodia theobromae contain messenger ribonucleic acid. Arch Biochem Biophys. 1970 Apr;137(2):442–452. doi: 10.1016/0003-9861(70)90461-3. [DOI] [PubMed] [Google Scholar]
  4. Chang L. M., Bollum F. J. Low molecular weight deoxyribonucleic acid polymerase from rabbit bone marrow. Biochemistry. 1972 Mar 28;11(7):1264–1272. doi: 10.1021/bi00757a023. [DOI] [PubMed] [Google Scholar]
  5. Crerar M., Pearlman R. E. DNA polymerase activity from Tetrahymena pyriformis. FEBS Lett. 1971 Nov 1;18(2):231–237. doi: 10.1016/0014-5793(71)80452-0. [DOI] [PubMed] [Google Scholar]
  6. Gong C. S., Van Etten J. L. Changes in soluble ribonucleic acid polymerases associated with the germination of Rhizopus stolonifer spores. Biochim Biophys Acta. 1972 Jun 22;272(1):44–52. doi: 10.1016/0005-2787(72)90031-7. [DOI] [PubMed] [Google Scholar]
  7. Howk R., Wang T. Y. DNA polymerase from rat liver chromosomal proteins. I. Partial purification and general characteristics. Arch Biochem Biophys. 1969 Sep;133(2):238–246. doi: 10.1016/0003-9861(69)90451-2. [DOI] [PubMed] [Google Scholar]
  8. Iwashima A., Rabinowitz M. Partial purification of mitochondrial and supernatant DNA polymerase from Saccharomyces cerevisiae. Biochim Biophys Acta. 1969 Apr 22;178(2):283–293. doi: 10.1016/0005-2744(69)90396-9. [DOI] [PubMed] [Google Scholar]
  9. Kalf G. F., Ch'ih J. J. Purification and properties of deoxyribonucleic acid polymerase from rat liver mitochondria. J Biol Chem. 1968 Sep 25;243(18):4904–4916. [PubMed] [Google Scholar]
  10. Klein A., Bonhoeffer F. DNA replication. Annu Rev Biochem. 1972;41(10):301–332. doi: 10.1146/annurev.bi.41.070172.001505. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. 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]
  13. Merlo D. J., Roker H., Van Etten J. L. Protein synthesis during fungal-spore germination. VI. Analysis of transfer ribonucleic acid from germinated and ungerminated spores of Rhizopus stolonifer. Can J Microbiol. 1972 Jul;18(7):949–956. doi: 10.1139/m72-147. [DOI] [PubMed] [Google Scholar]
  14. Meyer R. R., Simpson M. V. DNA biosynthesis in mitochondria: partial purification of a distinct DNA polymerase from isolated rat liver mitochondria. Proc Natl Acad Sci U S A. 1968 Sep;61(1):130–137. doi: 10.1073/pnas.61.1.130. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Schönherr O. T., Wanka F. An investigation of DNA polymerase in synchronously growing Chlorella cells. Biochim Biophys Acta. 1971 Feb 25;232(1):83–93. doi: 10.1016/0005-2787(71)90493-x. [DOI] [PubMed] [Google Scholar]
  17. Sedwick W. D., Wang T. S., Korn D. Purification and properties of nuclear and cytoplasmic deoxyribonucleic acid polymerases from human KB cells. J Biol Chem. 1972 Aug 25;247(16):5026–5033. [PubMed] [Google Scholar]
  18. Shapiro A. L., Viñuela E., Maizel J. V., Jr Molecular weight estimation of polypeptide chains by electrophoresis in SDS-polyacrylamide gels. Biochem Biophys Res Commun. 1967 Sep 7;28(5):815–820. doi: 10.1016/0006-291x(67)90391-9. [DOI] [PubMed] [Google Scholar]
  19. Stout E. R., Arens M. Q. DNA polymerase from maize seedlings. Biochim Biophys Acta. 1970 Jul 16;213(1):90–100. doi: 10.1016/0005-2787(70)90010-9. [DOI] [PubMed] [Google Scholar]
  20. Tarr H. L., Gardner L. Deoxyribonucleic acid polymerase from immature testes of salmon. Can J Biochem. 1971 Jan;49(1):19–27. doi: 10.1139/o71-004. [DOI] [PubMed] [Google Scholar]
  21. Van Etten J. L., Koski R. K., el-Olemy M. M. Protein synthesis during fungal spore germination. IV. Transfer ribonucleic acid from germinated and ungerminated spores. J Bacteriol. 1969 Dec;100(3):1182–1186. doi: 10.1128/jb.100.3.1182-1186.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Weissbach A., Schlabach A., Fridlender B., Bolden A. DNA polymerases from human cells. Nat New Biol. 1971 Jun 9;231(23):167–170. doi: 10.1038/newbio231167a0. [DOI] [PubMed] [Google Scholar]
  23. Wintersberger U., Wintersberger E. Studies on deoxyribonucleic acid polymerases from yeast. 1. Parial purification and properties of two DNA polymerases from mitochondria-free cell extracts. Eur J Biochem. 1970 Mar 1;13(1):11–19. doi: 10.1111/j.1432-1033.1970.tb00893.x. [DOI] [PubMed] [Google Scholar]
  24. 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]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES