Abstract
Chromatin isolated from adult rat liver was fractionated into template active and inactive components by controlled shearing and glycerol gradient centrifugation. The fractionated chromatin was assayed for DNA-dependent DNA polymerase (DNA mucleotidyl transferase EC 2.7.7.7) activity with and without exogenous activated DNA serving as template. With endogenous chromatin as template, it was found that 90% of the endogenous chromatin bound DNA polymerase activity was located in the transcriptionally active fraction of chromatin, while the distribution of DNA polymerase assayed with exogenous activated DNA was found to be 65% in the transcriptionally active and 35% in the inactive fractions. However, when DNA polymerase was solubilized from these fractions by salt extraction, enzyme activity was found to be equally distributed, suggesting that the difference in endogenous DNA polymerase activity observed between eu- and heterochromatin is due to the restricted template found in repressed fractions.
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Selected References
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- BURTON K. A study of the conditions and mechanism of the diphenylamine reaction for the colorimetric estimation of deoxyribonucleic acid. Biochem J. 1956 Feb;62(2):315–323. doi: 10.1042/bj0620315. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Baril E., Baril B., Laszlo J. DNA polymerases in normal and regenerating rat liver and hepatomas: targets for chemotherapy. Adv Enzyme Regul. 1974;12:355–372. doi: 10.1016/0065-2571(74)90021-1. [DOI] [PubMed] [Google Scholar]
- Bloch S., Cedar H. Methylation of chromatin DNA. Nucleic Acids Res. 1976 Jun;3(6):1507–1519. doi: 10.1093/nar/3.6.1507. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bollum F. J. Mammalian DNA polymerases. Prog Nucleic Acid Res Mol Biol. 1975;15(0):109–144. doi: 10.1016/s0079-6603(08)60118-x. [DOI] [PubMed] [Google Scholar]
- Chan J. Y., Srivastava B. I. Antigenic relationships in calf thymus and human leukemic cell terminal deoxynucleotidyl transferase. Biochim Biophys Acta. 1976 Oct 18;447(3):353–359. doi: 10.1016/0005-2787(76)90058-7. [DOI] [PubMed] [Google Scholar]
- Deumling B., Franke W. W. Nuclear membranes from mammalian liver, V. On the question of DNA polymerase activities associated with the nuclear envelope. Hoppe Seylers Z Physiol Chem. 1972 Mar;353(3):287–297. doi: 10.1515/bchm2.1972.353.1.287. [DOI] [PubMed] [Google Scholar]
- Doenecke D., McCarthy B. J. Movement of histones in chromatin induced by shearing. Eur J Biochem. 1976 May 1;64(2):405–409. doi: 10.1111/j.1432-1033.1976.tb10316.x. [DOI] [PubMed] [Google Scholar]
- FRENSTER J. H., ALLFREY V. G., MIRSKY A. E. REPRESSED AND ACTIVE CHROMATIN ISOLATED FROM INTERPHASE LYMPHOCYTES. Proc Natl Acad Sci U S A. 1963 Dec;50:1026–1032. doi: 10.1073/pnas.50.6.1026. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fansler B. S. Eukaryotic DNA polymerases: their association with the nucleus and relationship to DNA replication. Int Rev Cytol. 1974;Suppl 4:363–415. [PubMed] [Google Scholar]
- Frenster J. H. Localized strand separations within deoxyribonucleic acid during selective transcription. Nature. 1965 Nov 27;208(5013):894–896. doi: 10.1038/208894b0. [DOI] [PubMed] [Google Scholar]
- HAY E. D., REVEL J. P. The fine structure of the DNP component of the nucleus. An electron microscopic study utilizing autoradiography to localize DNA synthesis. J Cell Biol. 1963 Jan;16:29–51. doi: 10.1083/jcb.16.1.29. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Jalouzot R., Gontcharoff M., Briane D. Fractionation of plant chromatin with immobilized RNA-polymerase. FEBS Lett. 1975 Nov 15;59(2):221–224. doi: 10.1016/0014-5793(75)80379-6. [DOI] [PubMed] [Google Scholar]
- KLEVECZ R. R., HSU T. C. THE DIFFERENTIAL CAPACITY FOR RNA SYNTHESIS AMONG CHROMOSOMES: A CYTOLOGICAL APPROACH. Proc Natl Acad Sci U S A. 1964 Sep;52:811–817. doi: 10.1073/pnas.52.3.811. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lindahl T., Nyberg B. Rate of depurination of native deoxyribonucleic acid. Biochemistry. 1972 Sep 12;11(19):3610–3618. doi: 10.1021/bi00769a018. [DOI] [PubMed] [Google Scholar]
- Lindsay J. G., Berryman S., Adams R. L. Characteristics of deoxyribonucleic acid polymerase activity in nuclear and supernatant fractions of cultured mouse cells. Biochem J. 1970 Oct;119(5):839–848. doi: 10.1042/bj1190839. [DOI] [PMC free article] [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]
- Lynch W. E., Surrey S., Lieberman I. Nuclear deoxyribonucleic acid polymerases of liver. J Biol Chem. 1975 Oct 25;250(20):8179–8183. [PubMed] [Google Scholar]
- Montagna R. A., Rodriguez L. V., Becker F. F. A comparative study of the nonhistone proteins of rat liver euchromatin and heterochromatin. Arch Biochem Biophys. 1977 Mar;179(2):617–624. doi: 10.1016/0003-9861(77)90150-3. [DOI] [PubMed] [Google Scholar]
- Philippe M., Chevaillier P. Further characterization of a DNA polymerase activity in mouse sperm nuclei. Biochim Biophys Acta. 1976 Oct 4;447(2):188–202. doi: 10.1016/0005-2787(76)90342-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Philippe M., De Recondo A. M., Chevaillier P. Intranuclear distribution of mouse liver nuclear DNA polymerase. Exp Cell Res. 1976 Mar 15;98(2):424–428. doi: 10.1016/0014-4827(76)90451-1. [DOI] [PubMed] [Google Scholar]
- Rodriguez L. V., Becker F. F. Rat liver chromatin. Fractionation into eu- and heterochromatin with localization of ribosomal genes. Arch Biochem Biophys. 1976 Apr;173(2):428–437. doi: 10.1016/0003-9861(76)90280-0. [DOI] [PubMed] [Google Scholar]
- Srivastava B. I. Changes in enzymic activity during cultivation of human cells in vitro. Exp Cell Res. 1973 Aug;80(2):305–312. doi: 10.1016/0014-4827(73)90301-7. [DOI] [PubMed] [Google Scholar]
- Varshavskii A. I., Bakaev V. V., Il'in I. V., Baev A. A., Jr, Kadykov V. A. Struktura khromosomnykh dezoksiribonukleoproteidov. VII. Svobodnaia DNK v preparatakh fragmentirovannogo khromatina. Mol Biol (Mosk) 1976 Jan-Feb;10(1):35–54. [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]
