Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1978 Sep;27(3):576–581. doi: 10.1128/jvi.27.3.576-581.1978

Reverse transcriptase-associated RNase H activity. II. Inhibition by natural and synthetic RNA.

S L Marcus, S W Smith, M J Modak
PMCID: PMC525844  PMID: 81313

Abstract

The RNase H activity associated with purified avian myeloblastosis virus and Rauscher murine leukemia virus DNA polymerases is inhibited by homopolymeric RNA molecules, although the efficiency of inhibition by each homopolymer appears enzyme specific. Formation of duplex RNA molecules abolished the inhibitory activity. In contrast to these results, DNA polymerase-independent RNase H activities, such as the RNase H-II from Rauscher murine leukemia virus and calf thymus RNase H, were unaffected by the addition of exogenous RNA molecules to reaction mixtures. These results support the concept (M. J. Modak and S. L. Marcus, J. Virol. 22:253--256, 1977) that the catalytic site of DNA polymerase-associated RNase H activity may be that which is also involved in template binding. Naturally occurring RNA molecules of oncornaviral, procaryotic, or eucaryotic origin also proved to be efficient inhibitors of avian myeloblastosis virus DNA polymerase-associated RNase H activity. In contrast to this result, naturally occurring RNA molecules, at concentrations which inhibited the avian myeloblastosis virus enzyme, did not inhibit Rauscher murine leukemia virus DNA polymerase-catalyzed RNase H activity. This finding represents a new biochemical distinction between the two reverse transcriptases, and may be indicative of differences in the relative affinities of these enzymes for natural RNA molecules.

Full text

PDF
576

Selected References

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

  1. Abrell J. W., Smith R. G., Robert M. S., Gallo B. C. DNA polymerases from RNA tumor viruses and human cells: inhibition by polyuridylic acid. Science. 1972 Sep 22;177(4054):1111–1114. doi: 10.1126/science.177.4054.1111. [DOI] [PubMed] [Google Scholar]
  2. Brewer L. C., Wells R. D. Mechanistic independence of avian myeloblastosis virus DNA polymerase and ribonuclease H. J Virol. 1974 Dec;14(6):1494–1502. doi: 10.1128/jvi.14.6.1494-1502.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Erickson R. J., Grosch J. C. The inhibition of avian myeloblastosis virus deoxyribonucleic acid polymerase by synthetic polynucleotides. Biochemistry. 1974 Apr 23;13(9):1987–1993. doi: 10.1021/bi00706a032. [DOI] [PubMed] [Google Scholar]
  4. Erickson R. J., Janik B., Sommer R. G. The inhibition of the avian myeloblastosis virus DNA polymerase by poly(U) fractions of varying chain length. Biochem Biophys Res Commun. 1973 Jun 19;52(4):1475–1482. doi: 10.1016/0006-291x(73)90667-0. [DOI] [PubMed] [Google Scholar]
  5. Gerard G. F., Grandgenett D. P. Purification and characterization of the DNA polymerase and RNase H activities in Moloney murine sarcoma-leukemia virus. J Virol. 1975 Apr;15(4):785–797. doi: 10.1128/jvi.15.4.785-797.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. Marcus S. L., Modak M. J. Observations on template-specific conditions for DNA synthesis by avian myeloblastosis virus DNA polymerase. Nucleic Acids Res. 1976 Jun;3(6):1473–1486. doi: 10.1093/nar/3.6.1473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Marcus S. L. Resolution and characterization of intracytoplasmic forms of reverse transcriptase from Rauscher leukemia virus-producing cells. J Virol. 1978 Apr;26(1):1–10. doi: 10.1128/jvi.26.1.1-10.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Marcus S. L., Sarkar N. H., Modak M. J. Purification and properties of murine mammary tumor virus DNA polymerase. Virology. 1976 May;71(1):242–254. doi: 10.1016/0042-6822(76)90109-4. [DOI] [PubMed] [Google Scholar]
  10. Modak M. J., Marcus S. L. Purification and properties of Rauscher leukemia virus DNA polymerase and selective inhibition of mammalian viral reverse transcriptase by inorganic phosphate. J Biol Chem. 1977 Jan 10;252(1):11–19. [PubMed] [Google Scholar]
  11. Modak M. J. Pyridoxal 5' phosphate: a selective inhibitor of oncornaviral DNA polymerases. Biochem Biophys Res Commun. 1976 Jul 12;71(1):180–187. doi: 10.1016/0006-291x(76)90266-7. [DOI] [PubMed] [Google Scholar]
  12. Moelling K. Further characterization of the Friend murine leukemia virus reverse transcriptase-RNase H complex. J Virol. 1976 May;18(2):418–425. doi: 10.1128/jvi.18.2.418-425.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Stavrianopoulos J. G., Gambino-Giuffrida A., Chargaff E. Ribonuclease H of calf thymus: substrate specificity, activation, inhibition. Proc Natl Acad Sci U S A. 1976 Apr;73(4):1087–1091. doi: 10.1073/pnas.73.4.1087. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Verma I. M. Studies on reverse transcriptase of RNA tumor viruses III. Properties of purified Moloney murine leukemia virus DNA polymerase and associated RNase H. J Virol. 1975 Apr;15(4):843–854. doi: 10.1128/jvi.15.4.843-854.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Verma I. M. Studies on reverse transcriptase of RNA tumor viruses. I. Localization of thermolabile DNA polymerase and RNase H activities on one polypeptide. J Virol. 1975 Jan;15(1):121–126. doi: 10.1128/jvi.15.1.121-126.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES