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. 1974 Feb;71(2):578–582. doi: 10.1073/pnas.71.2.578

Multiple Forms of DNA Polymerase in Mouse Myeloma

A Matsukage 1, E W Bohn 1, S H Wilson 1
PMCID: PMC388051  PMID: 4521824

Abstract

Five distinct forms of DNA polymerase (deoxynucleosidetriphosphate: DNA deoxynucleotidyltransferase, EC 2.7.7.7) were separated from extracts of mouse myeloma MOPC-104E using a fractionation procedure based upon sequential ion-exchange column chromatography. The enzymes were characterized according to sedimentation behavior, subcellular localization, chromatographic behavior on hydroxyapatite columns, and reaction properties. The results indicate that myeloma contains two enzymes that appear to correspond to well characterized DNA polymerases found in many other mammalian tissues, a 6S DNA polymerase localized in the cytoplasmic supernatant fraction, and a lower molecular weight (2-3S) DNA polymerase. Also present were a second 6S DNA polymerase localized exclusively in the nuclear fraction and a 6-8S DNA polymerase localized in the cytoplasmic membrane fraction. The enzyme in the cytoplasmic membrane fraction, which accounted for the predominant activity in the myeloma, was active with poly(rA)·(dT)12-18 as template-primer, but not with activated calf thymus DNA. The detection of this distinct 6-8S membrane-bound DNA polymerase is of particular interest.

Keywords: systematic fractionation, poly(rA)·oligo(dT) template-primer, tumors

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Selected References

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  1. Baril E. F., Brown O. E., Jenkins M. D., Laszlo J. Deoxyribonucleic acid polymerase with rat liver ribosomes and smooth membranes. Purification and properties of the enzymes. Biochemistry. 1971 May 25;10(11):1981–1992. doi: 10.1021/bi00787a004. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Chang L. M., Bollum F. J. Low molecular weight deoxyribonucleic acid polymerase in mammalian cells. J Biol Chem. 1971 Sep 25;246(18):5835–5837. [PubMed] [Google Scholar]
  4. Fridlender B., Fry M., Bolden A., Weissbach A. A new synthetic RNA-dependent DNA polymerase from human tissue culture cells (HeLa-fibroblast-synthetic oligonucleotides-template-purified enzymes). Proc Natl Acad Sci U S A. 1972 Feb;69(2):452–455. doi: 10.1073/pnas.69.2.452. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Haines M. E., Holmes A. M., Johnston I. R. Distinct cytoplasmic and nuclear DNA polymerases from rat liver. FEBS Lett. 1971 Sep 15;17(1):63–67. doi: 10.1016/0014-5793(71)80564-1. [DOI] [PubMed] [Google Scholar]
  6. Iwamura Y., Ono T., Morris H. P. The heterogeneity of DNA polymerases in rat liver and hepatomas. Cancer Res. 1968 Dec;28(12):2466–2476. [PubMed] [Google Scholar]
  7. Maïa J. C.C., Rougeon F., Chapeville F. Chick embryo poly (rA:dT)-dependent DNA polymerase. FEBS Lett. 1971 Oct 15;18(1):130–134. doi: 10.1016/0014-5793(71)80427-1. [DOI] [PubMed] [Google Scholar]
  8. Ove P., Brown O. E., Laszlo J. Separation of DNA polymerase from rat liver and hepatomas. Cancer Res. 1969 Aug;29(8):1562–1567. [PubMed] [Google Scholar]
  9. Ove P., Coetzee M. L., Morris H. P. Separable DNA polymerase activities in host liver and Morris hepatomas. Cancer Res. 1973 Jun;33(6):1272–1283. [PubMed] [Google Scholar]
  10. Persico F. J., Gottlieb A. A. DNA polymerases of myeloma: template and primer specificities of two enzymes. Nat New Biol. 1972 Oct 11;239(93):173–176. doi: 10.1038/newbio239173a0. [DOI] [PubMed] [Google Scholar]
  11. Ross J., Scolnick E. M., Todaro G. J., Aaronson S. A. Separation of murine cellular and murine leukaemia virus DNA polymerases. Nat New Biol. 1971 Jun 9;231(23):163–167. doi: 10.1038/newbio231163a0. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Slater I., Slater D. W. DNA polymerase potentials of sea urchin embryos. Nat New Biol. 1972 May 17;237(72):81–85. doi: 10.1038/newbio237081a0. [DOI] [PubMed] [Google Scholar]
  14. Stavrianopoulos J. G., Karkas J. D., Chargaff E. Nucleic acid polymerases of the developing chicken embryo: a DNA polymerase preferring a hybrid template. Proc Natl Acad Sci U S A. 1971 Sep;68(9):2207–2211. doi: 10.1073/pnas.68.9.2207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Stromberg K., Gantt R., Wilson S. H. Structural studies on avian myeloblastosis virus: conditions for isolation and biochemical characteristics of the core component. Biochim Biophys Acta. 1973 Mar 30;304(1):1–11. doi: 10.1016/0304-4165(73)90109-8. [DOI] [PubMed] [Google Scholar]
  16. Suzuka I., Muto M. Presence of poly(rA):poly(dT)-dependent DNA polymerase in mouse myeloma cells. Biochem Biophys Res Commun. 1972 Mar 10;46(5):1874–1880. doi: 10.1016/0006-291x(72)90064-2. [DOI] [PubMed] [Google Scholar]
  17. Wallace P. G., Hewish D. R., Venning M. M., Burgoyne L. A. Multiple forms of mammalian deoxyribonucleic acid polymerase. An attempt to relate their interactions with nuclei and free deoxyribonucleic acid in vitro with their possible functions in vivo. Biochem J. 1971 Nov;125(1):47–54. doi: 10.1042/bj1250047. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Ward D. C., Humphryes K. C., Weinstein I. B. Synthetic RNA-dependent DNA polymerase activity in normal rat liver and hepatomas. Nature. 1972 Jun 30;237(5357):499–503. doi: 10.1038/237499a0. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Wilson S. H., Kuff E. L. A novel DNA polymerase activity found in association with intracisternal A-type particles. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1531–1536. doi: 10.1073/pnas.69.6.1531. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. YONEDA M., BOLLUM F. J. DEOXYNUCLEOTIDE-POLYMERIZING ENZYMES OF CALF THYMUS GLAND. I. LARGE SCALE PURIFICATION OF TERMINAL AND REPLICATIVE DEOXYNUCLEOTIDYL TRANSFERASES. J Biol Chem. 1965 Aug;240:3385–3391. [PubMed] [Google Scholar]
  22. Yoshida S., Modak M. J., Yagi K. DNA polymerase associated with nuclear membrane of calf thymus. Biochem Biophys Res Commun. 1971 Jun 18;43(6):1408–1414. doi: 10.1016/s0006-291x(71)80031-1. [DOI] [PubMed] [Google Scholar]

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