Abstract
Ribonuclease H of calf thymus has been purified better than 3000-fold to yield an almost homogeneous preparation. The enzyme, which comprises about 0.03% of the total protein in the initial extract, is a slightly acidic protein (pI = 4.95) of molecular weight of about 64,000, possibly composed of subunits. The enzyme requires a metal ion for activation; the conditions for activation by Mg, Co, and Mn are described. It is inhibited by S-adenosylmethionine. The substrates cleaved were poly(dT)·poly(rA) and the DNA-RNA hybrid made from phage f1 DNA; ribosomal RNA was not attacked.
Keywords: S-adenosylmethionine; Mg, Co, Mn; poly(dT)·poly(rA); phage f1 DNA
Full text
PDF
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Burgess R. R. A new method for the large scale purification of Escherichia coli deoxyribonucleic acid-dependent ribonucleic acid polymerase. J Biol Chem. 1969 Nov 25;244(22):6160–6167. [PubMed] [Google Scholar]
- Chang L. M., Bollum F. J. Deoxynucleotide-polymerizing enzymes of calf thymus gland. V. Homogeneous terminal deoxynucleotidyl transferase. J Biol Chem. 1971 Feb 25;246(4):909–916. [PubMed] [Google Scholar]
- Fairbanks G., Steck T. L., Wallach D. F. Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry. 1971 Jun 22;10(13):2606–2617. doi: 10.1021/bi00789a030. [DOI] [PubMed] [Google Scholar]
- Karkas J. D. Coupling of replication to transcription in vitro. Proc Natl Acad Sci U S A. 1972 Aug;69(8):2288–2291. doi: 10.1073/pnas.69.8.2288. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Karkas J. D., Stavrianopoulos J. G., Chargaff E. Action of DNA polymerase I of Escherichia coli with DNA-RNA hybrids as templates. Proc Natl Acad Sci U S A. 1972 Feb;69(2):398–402. doi: 10.1073/pnas.69.2.398. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Keller W., Crouch R. Degradation of DNA RNA hybrids by ribonuclease H and DNA polymerases of cellular and viral origin. Proc Natl Acad Sci U S A. 1972 Nov;69(11):3360–3364. doi: 10.1073/pnas.69.11.3360. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Keller W. RNA-primed DNA synthesis in vitro. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1560–1564. doi: 10.1073/pnas.69.6.1560. [DOI] [PMC free article] [PubMed] [Google Scholar]
- POLSON A., POTGIETER G. M., LARGIER J. F., MEARS G. E., JOUBERT F. J. THE FRACTIONATION OF PROTEIN MIXTURES BY LINEAR POLYMERS OF HIGH MOLECULAR WEIGHT. Biochim Biophys Acta. 1964 Mar 16;82:463–475. doi: 10.1016/0304-4165(64)90438-6. [DOI] [PubMed] [Google Scholar]
- Stavrianopoulos J. G., Karkas J. D., Chargaff E. DNA polymerase of chicken embryo: purification and properties. Proc Natl Acad Sci U S A. 1972 Jul;69(7):1781–1785. doi: 10.1073/pnas.69.7.1781. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stavrianopoulos J. G., Karkas J. D., Chargaff E. Mechanism of DNA replication by highly purified DNA polymerase of chicken embryo. Proc Natl Acad Sci U S A. 1972 Sep;69(9):2609–2613. doi: 10.1073/pnas.69.9.2609. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Stein H., Hausen P. Enzyme from calf thymus degrading the RNA moiety of DNA-RNA Hybrids: effect on DNA-dependent RNA polymerase. Science. 1969 Oct 17;166(3903):393–395. doi: 10.1126/science.166.3903.393. [DOI] [PubMed] [Google Scholar]