Abstract
In vitro DNA synthesis on single-stranded circular DNA can be initiated by RNA primers. RNA chains are covalently extended by DNA polymerase II from KB cells and DNA polymerase I from Micrococcus luteus, but not by an RNA-dependent DNA polymerase from avian myeloblastosis virus. The reaction product consists of DNA chains with a piece of RNA at their 5′-ends, hydrogen bonded to the template DNA. The primer RNA is linked to the product DNA via a 3′:5′-phosphodiester bond, and can be specifically removed by ribonuclease H. The possible role of ribonuclease H in RNA-primed DNA synthesis in vivo is discussed.
Keywords: DNA polymerase, DNA initiation, RNA polymerase, ribonuclease H
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Selected References
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- Bazzicalupo P., Tocchini-Valentini G. P. Curing of an Escherichia coli episome by rifampicin (acridine orange-F + -F - -Hfr-lac). Proc Natl Acad Sci U S A. 1972 Feb;69(2):298–300. doi: 10.1073/pnas.69.2.298. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Brutlag D., Schekman R., Kornberg A. A possible role for RNA polymerase in the initiation of M13 DNA synthesis. Proc Natl Acad Sci U S A. 1971 Nov;68(11):2826–2829. doi: 10.1073/pnas.68.11.2826. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- CHAMBERLIN M., BERG P. MECHANISM OF RNA POLYMERASE ACTION: FORMATION OF DNA-RNA HYBRIDS WITH SINGLE-STRANDED TEMPLATES. J Mol Biol. 1964 Feb;8:297–313. doi: 10.1016/s0022-2836(64)80139-x. [DOI] [PubMed] [Google Scholar]
- Fanshier L., Garapin A. C., McDonnell J., Faras A., Levinson W., Bishop J. M. Deoxyribonucleic acid polymerase associated with avian tumor viruses: secondary structure of the deoxyribonucleic acid product. J Virol. 1971 Jan;7(1):77–86. doi: 10.1128/jvi.7.1.77-86.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hausen P., Stein H. Ribonuclease H. An enzyme degrading the RNA moiety of DNA-RNA hybrids. Eur J Biochem. 1970 Jun;14(2):278–283. doi: 10.1111/j.1432-1033.1970.tb00287.x. [DOI] [PubMed] [Google Scholar]
- Hurwitz J., Leis J. P. RNA-dependent DNA polymerase activity of RNA tumor viruses. I. Directing influence of DNA in the reaction. J Virol. 1972 Jan;9(1):116–129. doi: 10.1128/jvi.9.1.116-129.1972. [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]
- Lark K. G. Evidence for the direct involvement of RNA in the initiation of DNA replication in Escherichia coli 15T. J Mol Biol. 1972 Feb 28;64(1):47–60. doi: 10.1016/0022-2836(72)90320-8. [DOI] [PubMed] [Google Scholar]
- Leis J. P., Hurwitz J. RNA-dependent DNA polymerase activity of RNA tumor viruses. II. Directing influence of RNA in the reaction. J Virol. 1972 Jan;9(1):130–142. doi: 10.1128/jvi.9.1.130-142.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Manly K. F., Smoler D. F., Bromfeld E., Baltimore D. Forms of deoxyribonucleic acid produced by virions of the ribonucleic acid tumor viruses. J Virol. 1971 Jan;7(1):106–111. doi: 10.1128/jvi.7.1.106-111.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mölling K., Bolognesi D. P., Bauer H., Büsen W., Plassmann H. W., Hausen P. Association of viral reverse transcriptase with an enzyme degrading the RNA moiety of RNA-DNA hybrids. Nat New Biol. 1971 Dec 22;234(51):240–243. doi: 10.1038/newbio234240a0. [DOI] [PubMed] [Google Scholar]
- Rabin E. Z., Fraser M. J. Isolation of Neurospora crassa endonuclease specific for single-stranded DNA. Can J Biochem. 1970 Mar;48(3):389–392. doi: 10.1139/o70-063. [DOI] [PubMed] [Google Scholar]
- Robertson H. D., Webster R. E., Zinder N. D. Purification and properties of ribonuclease III from Escherichia coli. J Biol Chem. 1968 Jan 10;243(1):82–91. [PubMed] [Google Scholar]
- Sedat J., Lyon A., Sinsheimer R. L. Purification of Escherichia coli pulse-labeled RNA by benzoylated DEAE-cellulose chromatography. J Mol Biol. 1969 Sep 28;44(3):415–434. doi: 10.1016/0022-2836(69)90370-2. [DOI] [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]
- Verma I. M., Meuth N. L., Bromfeld E., Manly K. F., Baltimore D. Covalently linked RNA-DNA molecule as initial product of RNA tumour virus DNA polymerase. Nat New Biol. 1971 Sep 29;233(39):131–134. doi: 10.1038/newbio233131a0. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- Zimmerman B. K. Purification and properties of deoxyribonucleic acid polymerase from Micrococcus lysodeikticus. J Biol Chem. 1966 May 10;241(9):2035–2041. [PubMed] [Google Scholar]