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. 1978 Nov;75(11):5329–5333. doi: 10.1073/pnas.75.11.5329

Sodium pyrophosphate inhibition of RNA.DNA hybrid degradation by reverse transcriptase.

J C Myers, S Spiegelman
PMCID: PMC392956  PMID: 82966

Abstract

Sodium pyrophosphate inhibits synthesis of anticomplementary DNA during a reverse transcriptase (RNA-directed DNA nucleotidyltransferase, EC 2.7.7.7) catalyzed reaction. In the presence of pyrophosphate, the complementary DNA remains stably complexed to the RNA template. In the absence of pyrophosphate, the DNA. RNA hybrid template is degraded and anticomplementary DNA is synthesized. High concentrations of additives containing phosphodiester bonds appear to inhibit the ribonuclease H activity (hybrid nuclease, EC 3.1.4.34) of the reverse transcriptase, therby preventing formation of RNA primers necessary for the synthesis of anticomplementary DNA.

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Selected References

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

  1. Dube D. K., Loeb L. A. On the association of reverse transcriptase with polynucleotide templates during catalysis. Biochemistry. 1976 Aug 10;15(16):3605–3611. doi: 10.1021/bi00661a031. [DOI] [PubMed] [Google Scholar]
  2. Efstratiadis A., Maniatis T., Kafatos F. C., Jeffrey A., Vournakis J. N. Full length and discrete partial reverse transcripts of globin and chorion mRNAs. Cell. 1975 Apr;4(4):367–378. doi: 10.1016/0092-8674(75)90157-9. [DOI] [PubMed] [Google Scholar]
  3. Faust C. H., Jr, Diggelmann H., Mach B. Isolation of poly(adenylic acid)-rich ribonucleic acid from mouse myeloma and synthesis of complementary deoxyribonucleic acid. Biochemistry. 1973 Feb 27;12(5):925–931. doi: 10.1021/bi00729a021. [DOI] [PubMed] [Google Scholar]
  4. Friedman E. Y., Rosbash M. The syntheiss of high yields of full-length reverse transcripts of globin mRNA. Nucleic Acids Res. 1977 Oct;4(10):3455–3471. doi: 10.1093/nar/4.10.3455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Imaizumi T., Diggelmann H., Scherrer K. Demonstration of globin messenger sequences in giant nuclear precursors of messenger RNA of avian erythroblasts. Proc Natl Acad Sci U S A. 1973 Apr;70(4):1122–1126. doi: 10.1073/pnas.70.4.1122. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Kacian D. L., Myers J. C. Anticomplementary nature of smaller DNA produced during synthesis of extensive DNA copies of poliovirus RNA. Proc Natl Acad Sci U S A. 1976 Oct;73(10):3408–3412. doi: 10.1073/pnas.73.10.3408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Kacian D. L., Myers J. C. Synthesis of extensive, possibly complete, DNA copies of poliovirus RNA in high yields and at high specific activities. Proc Natl Acad Sci U S A. 1976 Jul;73(7):2191–2195. doi: 10.1073/pnas.73.7.2191. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Kacian D. L., Spiegelman S. Purification and detection of reverse transcriptase in viruses and cells. Methods Enzymol. 1974;29:150–173. doi: 10.1016/0076-6879(74)29018-9. [DOI] [PubMed] [Google Scholar]
  9. Kacian D. L., Spiegelman S. Use of micrococcal nuclease to monitor hybridization reactions with DNA. Anal Biochem. 1974 Apr;58(2):534–540. doi: 10.1016/0003-2697(74)90221-8. [DOI] [PubMed] [Google Scholar]
  10. Leis J. P., Berkower I., Hurwitz J. Mechanism of action of ribonuclease H isolated from avian myeloblastosis virus and Escherichia coli. Proc Natl Acad Sci U S A. 1973 Feb;70(2):466–470. doi: 10.1073/pnas.70.2.466. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Leis J. P. RNA-dependent DNA polymerase activity of RNA tumor virus. VI. Processive mode of action of avian myeloblastosis virus polymerase. J Virol. 1976 Sep;19(3):932–939. doi: 10.1128/jvi.19.3.932-939.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Monahan J. J., Harris S. E., Woo S. L., Robberson D. L., O'Malley B. W. The synthesis and properties of the complete complementary DNA transcript of ovalbumin mRNA. Biochemistry. 1976 Jan 13;15(1):223–233. doi: 10.1021/bi00646a034. [DOI] [PubMed] [Google Scholar]
  13. Myers J. C., Spiegelman S., Kacian D. L. Synthesis of full-length DNA copies of avian myeloblastosis virus RNA in high yields. Proc Natl Acad Sci U S A. 1977 Jul;74(7):2840–2843. doi: 10.1073/pnas.74.7.2840. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Rothenberg E., Baltimore D. Synthesis of long, representative DNA copies of the murine RNA tumor virus genome. J Virol. 1975 Jan;17(1):168–174. doi: 10.1128/jvi.17.1.168-174.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Shank P. R., Varmus H. E. Virus-specific DNA in the cytoplasm of avian sarcoma virus-infected cells is a precursor to covalently closed circular viral DNA in the nucleus. J Virol. 1978 Jan;25(1):104–104. doi: 10.1128/jvi.25.1.104-104.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]

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