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. 1981 May;78(5):3049–3052. doi: 10.1073/pnas.78.5.3049

Intracellular localization of DNA polymerase alpha.

M Brown, F J Bollum, L M Chang
PMCID: PMC319497  PMID: 7019918

Abstract

Immunoglobulin (IgG) and the F(ab')2 fragment of IgG were prepared from serum of a rabbit immunized with purified calf thymus DNA polymerase alpha (deoxynucleosidetriphosphate:DNA deoxynucleotidyltransferase, EC 2.7.7.7). An indirect immunofluorescent method based on these reagents was used to detect the intracellular localization of DNA polymerase alpha in primary fetal bovine fibroblasts. The results show that the bulk of DNA polymerase alpha is located in the perinuclear region of the cytoplasm. Immunofluorescent staining of cytoplast and Ficoll-Paque gradient-purified karyoplast fragments resulting from cytochalasin enucleation show the presence of DNA polymerase alpha in cytoplasts and the virtual absence of the enzyme in the nucleus of the karyoplast itself. The implication of this unusual intracellular location for DNA polymerase alpha is discussed.

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Selected References

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

  1. BOLLUM F. J., POTTER V. R. Incorporation of thymidine into deoxyribonucleic acid by enzymes from rat tissues. J Biol Chem. 1958 Aug;233(2):478–482. [PubMed] [Google Scholar]
  2. Bertazzoni U., Scovassi A. I., Brun G. M. Chick-embryo DNA polymerase gamma. Identity of gamma-polymerases purified from nuclei and mitochondria. Eur J Biochem. 1977 Dec 1;81(2):237–248. doi: 10.1111/j.1432-1033.1977.tb11945.x. [DOI] [PubMed] [Google Scholar]
  3. Chang L. M., Bollum F. J. Immunological reagents for comparisons of DNA polymerase-alpha and DNA polymerase-beta. J Biol Chem. 1981 Jan 10;256(1):494–498. [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. Chang L. M., Bollum F. J. Variation of deoxyribonucleic acid polymerase activities during rat liver regeneration. J Biol Chem. 1972 Dec 25;247(24):7948–7950. [PubMed] [Google Scholar]
  6. 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]
  7. Foster D. N., Gurney T., Jr Nuclear location of mammalian DNA polymerase activities. J Biol Chem. 1976 Dec 25;251(24):7893–7898. [PubMed] [Google Scholar]
  8. Herrick G., Spear B. B., Veomett G. Intracellular localization of mouse DNA polymerase-alpha. Proc Natl Acad Sci U S A. 1976 Apr;73(4):1136–1139. doi: 10.1073/pnas.73.4.1136. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hübscher U., Kuenzle C. C., Spadari S. Identity of DNA polymerase gamma from synaptosomal mitochondria and rat-brain nuclei. Eur J Biochem. 1977 Dec 1;81(2):249–258. doi: 10.1111/j.1432-1033.1977.tb11946.x. [DOI] [PubMed] [Google Scholar]
  10. KEIR H. M., SMELLIE R. M., SIEBERT G. Intracellular location of DNA nucleotidyltransferase. Nature. 1962 Nov 24;196:752–754. doi: 10.1038/196752a0. [DOI] [PubMed] [Google Scholar]
  11. Keir H. M. DNA polymerases from mammalian cells. Prog Nucleic Acid Res Mol Biol. 1965;4:81–128. doi: 10.1016/s0079-6603(08)60785-0. [DOI] [PubMed] [Google Scholar]
  12. LITTLEFIELD J. W., McGOVERN A. P., MARGESON K. B. Changes in the distribution of polymerase activity during DNA synthesis in mouse fibroblasts. Proc Natl Acad Sci U S A. 1963 Jan 15;49:102–107. doi: 10.1073/pnas.49.1.102. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lucas J. J., Szekely E., Kates J. R. The regeneration and division of mouse L-cell karyoplasts. Cell. 1976 Jan;7(1):115–122. doi: 10.1016/0092-8674(76)90261-0. [DOI] [PubMed] [Google Scholar]
  14. PRESCOTT D. M., BOLLUM F. J., KLUSS B. C. Is DNA polymerase a cytoplasmic enzyme? J Cell Biol. 1962 Apr;13:172–174. doi: 10.1083/jcb.13.1.172. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Prem veer Reddy G., Pardee A. B. Multienzyme complex for metabolic channeling in mammalian DNA replication. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3312–3316. doi: 10.1073/pnas.77.6.3312. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Prescott D. M., Myerson D., Wallace J. Enucleation of mammalian cells with cytochalasin B. Exp Cell Res. 1972;71(2):480–485. doi: 10.1016/0014-4827(72)90322-9. [DOI] [PubMed] [Google Scholar]
  17. Shay J. W., Porter K. R., Prescott D. M. The surface morphology and fine structure of CHO (Chinese hamster ovary) cells following enucleation. Proc Natl Acad Sci U S A. 1974 Aug;71(8):3059–3063. doi: 10.1073/pnas.71.8.3059. [DOI] [PMC free article] [PubMed] [Google Scholar]

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