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. 1982 Apr 10;10(7):2309–2321. doi: 10.1093/nar/10.7.2309

An oligoribonucleotide polymerase from SV40-infected cells with properties of a primase.

G Kaufmann, H H Falk
PMCID: PMC320611  PMID: 6283476

Abstract

A transient decaribonucleotide (iRNA) is covalently linked to nascent eukaryotic DNA chains at their 5' end. Searching for the putative iRNA polymerase (primase), we detected in extracts from SV40-infected cells a DNA-dependent incorporation of UMP residues from UTP into free and DNA linked deca- or similarly sized ribonucleotides. Denatured salmon sperm DNA served as the standard template in this reaction. SV40 FIII DNA was also an effective template, SV40 FII DNA was ineffective while FI yielded mainly free decaribonucleotides. The incorporation depended on the other rNTPs and was resistant to high concentrations of alpha-amanitin and rifamycin AF/013, drugs inhibitory to RNA polymerases I, II and III. The results implicate the decaribonucleotide polymerase in the priming of nascent DNA chains and suggest that the unique size of iRNA is encoded within its primase.

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Selected References

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

  1. Bouché J. P., Zechel K., Kornberg A. dnaG gene product, a rifampicin-resistant RNA polymerase, initiates the conversion of a single-stranded coliphage DNA to its duplex replicative form. J Biol Chem. 1975 Aug 10;250(15):5995–6001. [PubMed] [Google Scholar]
  2. Champoux J. J., McConaughy B. L. Priming of superhelical SV40 DNA by Escherichia coli RNA polymerase for in vitro DNA synthesis. Biochemistry. 1975 Jan 28;14(2):307–316. doi: 10.1021/bi00673a017. [DOI] [PubMed] [Google Scholar]
  3. Eliasson R., Reichard P. Primase initiates Okazaki pieces during polyoma DNA synthesis. Nature. 1978 Mar 9;272(5649):184–185. doi: 10.1038/272184a0. [DOI] [PubMed] [Google Scholar]
  4. Eliasson R., Reichard P. Replication of polyoma DNA in isolated nuclei. VII. Initiator RNA synthesis during nucleotide depletion. J Mol Biol. 1979 Apr 15;129(3):393–409. doi: 10.1016/0022-2836(79)90503-5. [DOI] [PubMed] [Google Scholar]
  5. Hunter T., Francke B. In vitro polyoma DNA synthesis: characterization of a system from infected 3T3 cells. J Virol. 1974 Jan;13(1):125–139. doi: 10.1128/jvi.13.1.125-139.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Kaufmann G., Anderson S., DePamphilis M. L. RNA primers in Simian virus 40 DNA replication. II. Distribution of 5' terminal oligoribonucleotides in nascent DNA. J Mol Biol. 1977 Nov 5;116(3):549–567. doi: 10.1016/0022-2836(77)90083-3. [DOI] [PubMed] [Google Scholar]
  7. Kaufmann G. Characterization of initiator RNA from replicating simian virus 40 DNA synthesized in isolated nuclei. J Mol Biol. 1981 Mar 25;147(1):25–39. doi: 10.1016/0022-2836(81)90077-2. [DOI] [PubMed] [Google Scholar]
  8. Kedinger C., Gniazdowski M., Mandel J. L., Jr, Gissinger F., Chambon P. Alpha-amanitin: a specific inhibitor of one of two DNA-pendent RNA polymerase activities from calf thymus. Biochem Biophys Res Commun. 1970 Jan 6;38(1):165–171. doi: 10.1016/0006-291x(70)91099-5. [DOI] [PubMed] [Google Scholar]
  9. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  10. Magnusson G., Pigiet V., Winnacker E. L., Abrams R., Reichard P. RNA-linked short DNA fragments during polyoma replication. Proc Natl Acad Sci U S A. 1973 Feb;70(2):412–415. doi: 10.1073/pnas.70.2.412. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Manley J. L., Fire A., Cano A., Sharp P. A., Gefter M. L. DNA-dependent transcription of adenovirus genes in a soluble whole-cell extract. Proc Natl Acad Sci U S A. 1980 Jul;77(7):3855–3859. doi: 10.1073/pnas.77.7.3855. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Meilhac M., Tysper Z., Chambon P. Animal DNA-dependent RNA polymerases. 4. Studies on inhibition by rifamycin derivatives. Eur J Biochem. 1972 Jul 13;28(2):291–300. doi: 10.1111/j.1432-1033.1972.tb01913.x. [DOI] [PubMed] [Google Scholar]
  13. Noll M. Internal structure of the chromatin subunit. Nucleic Acids Res. 1974 Nov;1(11):1573–1578. doi: 10.1093/nar/1.11.1573. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. RAZZELL W. E., KHORANA H. G. Studies on polynucleotides. III. Enzymic degradation; substrate specificity and properties of snake venom phosphodiesterase. J Biol Chem. 1959 Aug;234(8):2105–2113. [PubMed] [Google Scholar]
  15. Reichard P., Eliasson R., Söderman G. Initiator RNA in discontinuous polyoma DNA synthesis. Proc Natl Acad Sci U S A. 1974 Dec;71(12):4901–4905. doi: 10.1073/pnas.71.12.4901. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Roeder R. G., Rutter W. J. Multiple forms of DNA-dependent RNA polymerase in eukaryotic organisms. Nature. 1969 Oct 18;224(5216):234–237. doi: 10.1038/224234a0. [DOI] [PubMed] [Google Scholar]
  17. Scherzinger E., Lanka E., Morelli G., Seiffert D., Yuki A. Bacteriophage-T7-induced DNA-priming protein. A novel enzyme involved in DNA replication. Eur J Biochem. 1977 Feb;72(3):543–558. doi: 10.1111/j.1432-1033.1977.tb11278.x. [DOI] [PubMed] [Google Scholar]
  18. Tseng B. Y., Goulian M. Initiator RNA of discontinuous DNA synthesis in human lymphocytes. Cell. 1977 Oct;12(2):483–489. doi: 10.1016/0092-8674(77)90124-6. [DOI] [PubMed] [Google Scholar]
  19. Tseng B. Y., Goulian M. Initiator RNA synthesis upon ribonucleotide depletion. Evidence for base substitutions. J Biol Chem. 1980 Mar 10;255(5):2062–2066. [PubMed] [Google Scholar]
  20. Weil P. A., Blatti S. P. Partial purification and properties of calf thymus deoxyribonucleic acid dependent RNA polymerase III. Biochemistry. 1975 Apr 22;14(8):1636–1642. doi: 10.1021/bi00679a015. [DOI] [PubMed] [Google Scholar]
  21. Weinmann R., Roeder R. G. Role of DNA-dependent RNA polymerase 3 in the transcription of the tRNA and 5S RNA genes. Proc Natl Acad Sci U S A. 1974 May;71(5):1790–1794. doi: 10.1073/pnas.71.5.1790. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Yagura T., Kozu T., Seno T. Mouse DNA polymerase accompanied by a novel RNA polymerase activity: purification and partial characterization. J Biochem. 1982 Feb;91(2):607–618. doi: 10.1093/oxfordjournals.jbchem.a133732. [DOI] [PubMed] [Google Scholar]

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