Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1969 May;112(5):721–727. doi: 10.1042/bj1120721

Experimental conditions affecting ribonucleic acid polymerase in isolated rat liver nuclei. Effect of nucleoside triphosphate concentration, temperature, ammonium sulphate and heparin

F Novello 1, F Stirpe 1
PMCID: PMC1187776  PMID: 5821730

Abstract

1. The conditions affecting the activity of RNA polymerase in isolated rat liver nuclei were studied with Mg2+ or Mn2+ as activating ions. 2. The enzyme assayed with Mg2+ and at low ionic strength is saturated by a lower concentration of nucleotide substrates than if assayed with Mn2+ at low ionic strength or with either ion at high ionic strength. 3. At low and at high ionic strength the incorporation of AMP is affected in a similar way by variations in the temperature of incubation. Preincubation at 37° impairs the AMP incorporation. 4. Heparin stimulates the RNA polymerase activity in the presence of Mn2+. 5. Both ammonium sulphate and heparin `restart' the reaction if added after 15min., the effect being more marked with ammonium sulphate than with heparin, and also more marked in the presence of Mn2+ than of Mg2+. 6. α-Amanitin abolishes the effect of ammonium sulphate and of heparin.

Full text

PDF
723

Selected References

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

  1. BALLARD P., WILLIAMS-ASHMAN H. G. ISOLATION OF A SOLUBLE RNA POLYMERASE FROM RAT TESTIS. Nature. 1964 Jul 11;203:150–151. doi: 10.1038/203150a0. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Bannai S., Terayama H. Regulation of RNA synthesis in chromatin (aggregate enzyme) from rat liver. Biochim Biophys Acta. 1967 Jul 18;142(2):410–418. doi: 10.1016/0005-2787(67)90622-3. [DOI] [PubMed] [Google Scholar]
  4. Barnabei O., Romano B., Di Bitonto G., Tomasi V., Sereni F. Factors influencing the glucocorticoid-induced increase of ribonucleic acid polymerase activity of rat liver nuclei. Arch Biochem Biophys. 1966 Feb;113(2):478–486. doi: 10.1016/0003-9861(66)90217-7. [DOI] [PubMed] [Google Scholar]
  5. Bresnick E. Ribonucleic acid polymerase activity in liver nuclei from rats pretreated with 3-methylcholanthrene. Mol Pharmacol. 1966 Sep;2(5):406–410. [PubMed] [Google Scholar]
  6. DAVIES H. G., SPENCER M. The variation in the structure of erythrocyte nuclei with fixation. J Cell Biol. 1962 Sep;14:445–458. doi: 10.1083/jcb.14.3.445. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. GOLDBERG I. H. Ribonucleic acid synthesis in nuclear extracts of mammalian cells grown in suspension culture; effect of ionic strength and surface-active agents. Biochim Biophys Acta. 1961 Jul 22;51:201–204. doi: 10.1016/0006-3002(61)91042-3. [DOI] [PubMed] [Google Scholar]
  8. Homoki J., Lukács I., Sekeris C. E. Effect of ammonium sulfate and other salts on the ribonuclease activity of isolated rat liver nuclei. Hoppe Seylers Z Physiol Chem. 1967 Nov;348(11):1392–1394. doi: 10.1515/bchm2.1967.348.1.1392. [DOI] [PubMed] [Google Scholar]
  9. Jacob S. T., Sajdel E. M., Munro H. N. Altered characteristics of mammalian RNA polymerase following solubilization from nuclei. Biochem Biophys Res Commun. 1968 Sep 6;32(5):831–838. doi: 10.1016/0006-291x(68)90316-1. [DOI] [PubMed] [Google Scholar]
  10. KENT P. W., HICHENS M., WARD P. F. Displacement fractionation of deoxyribonucleoproteins by heparin and dextran sulphate. Biochem J. 1958 Apr;68(4):568–572. doi: 10.1042/bj0680568. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Korohoda W., Forrester J. A., Moreman K. G., Ambrose E. J. Size changes in isolated nuclei of Amoeba proteus on treatment with polyionic substances. Nature. 1968 Feb 17;217(5129):615–617. doi: 10.1038/217615a0. [DOI] [PubMed] [Google Scholar]
  12. MacGregor R. R., Mahler H. R. RNA synthesis in intact rat liver nuclei. Arch Biochem Biophys. 1967 Apr;120(1):136–157. doi: 10.1016/0003-9861(67)90607-8. [DOI] [PubMed] [Google Scholar]
  13. Marchok A. C., Wolff J. A. Studies of muscle development. IV. Some characteristics of RNA polymerase activity in isolated nuclei from developing chick muscle. Biochim Biophys Acta. 1968 Feb 26;155(2):378–393. doi: 10.1016/0005-2787(68)90182-2. [DOI] [PubMed] [Google Scholar]
  14. Pogo B. G., Allfrey V. G., Mirsky A. E. RNA synthesis and histone acetylation during the course of gene activation in lymphocytes. Proc Natl Acad Sci U S A. 1966 Apr;55(4):805–812. doi: 10.1073/pnas.55.4.805. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Steiner D. F., King J. Insulin-stimulated ribonucleic acid synthesis and RNA polymerase activity in alloxan-diabetic rat liver. Biochim Biophys Acta. 1966 Jun 22;119(3):510–516. doi: 10.1016/0005-2787(66)90127-4. [DOI] [PubMed] [Google Scholar]
  16. Stirpe F., Fiume L. Studies on the pathogenesis of liver necrosis by alpha-amanitin. Effect of alpha-amanitin on ribonucleic acid synthesis and on ribonucleic acid polymerase in mouse liver nuclei. Biochem J. 1967 Nov;105(2):779–782. doi: 10.1042/bj1050779. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. WIDNELL C. C., TATA J. R. EVIDENCE FOR TWO DNA-DEPENDENT RNA POLYMERASE ACTIVITIES IN ISOLATED RAT-LIVER NUCLEI. Biochim Biophys Acta. 1964 Jul 22;87:531–533. doi: 10.1016/0926-6550(64)90133-1. [DOI] [PubMed] [Google Scholar]
  18. Walter G., Zillig W., Palm P., Fuchs E. Initiation of DNA-dependent RNA synthesis and the effect of heparin on RNA polymerase. Eur J Biochem. 1967 Dec;3(2):194–201. doi: 10.1111/j.1432-1033.1967.tb19515.x. [DOI] [PubMed] [Google Scholar]
  19. Widnell C. C., Tata J. R. A procedure for the isolation of enzymically active rat-liver nuclei. Biochem J. 1964 Aug;92(2):313–317. doi: 10.1042/bj0920313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Widnell C. C., Tata J. R. Studies on the stimulation by ammonium sulphate of the DNA-dependent RNA polymerase of isolated rat-liver nuclei. Biochim Biophys Acta. 1966 Sep;123(3):478–492. doi: 10.1016/0005-2787(66)90216-4. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES