Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1975 Oct;16(4):797–806. doi: 10.1128/jvi.16.4.797-806.1975

Purification and properties of spleen necrosis virus DNA polymerase.

A Mizutani, H M Temin
PMCID: PMC354738  PMID: 51934

Abstract

DNA polymerase was purified to apparent electrophoretic homogeneity from virions of spleen necrosis virus (SNV). (SNV is a member of the reticuloendotheliosis group of avian ribodeoxyviruses). The SNV DNA polymerase appears to consist of a single polypeptide with a molecular weight of 68,000. The SNV DNA polymerase has a preference for Mn2+ for DNA synthesis with an RNA template and Mg2+ for DNA synthesis with a deoxyribohomopolymer template. At the optimum concentrations of divalent cation, the relative rates of DNA synthesis by SNV DNA polymerase with different template.primers were similar to the relative rates of DNA synthesis by an avian leukosis virus DNA polymerase, with the exception of a lower relative rate of DNA synthesis by SNV DNA polymerase with SNV RNA. However, in contrast to DNA synthesized by the avian leukosis virus DNA polymerase with a SNV RNA template, DNA synthesized by SNV DNA polymerase with an SNV RNA template did not hybridize to the SNV RNA. SNV DNA polymerase has RNase H activity which is antigenically distinct from the RNase H activity of avian leukosis-sarcoma virus DNA polymerase.

Full text

PDF
797

Images in this article

800Image
on p.800

Selected References

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

  1. Cooper G. M., Temin H. M. Infectious rous sarcoma virus and reticuloendotheliosis virus DNAs. J Virol. 1974 Nov;14(5):1132–1141. doi: 10.1128/jvi.14.5.1132-1141.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Gibson W., Verma I. M. Studies on the reverse transcriptase of RNA tumor viruses. Structural relatedness of two subunits of avian RNA tumor viruses. Proc Natl Acad Sci U S A. 1974 Dec;71(12):4991–4994. doi: 10.1073/pnas.71.12.4991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Grandgenett D. P., Gerard G. F., Green M. A single subunit from avian myeloblastosis virus with both RNA-directed DNA polymerase and ribonuclease H activity. Proc Natl Acad Sci U S A. 1973 Jan;70(1):230–234. doi: 10.1073/pnas.70.1.230. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Grandgenett D. P., Green M. Different mode of action of ribonuclease H in purified alpha and alpha beta ribonucleic acid-directed deoxyribonucleic acid polymerase from avian myeloblastosis virus. J Biol Chem. 1974 Aug 25;249(16):5148–5152. [PubMed] [Google Scholar]
  5. Green M., Gerard G. F. RNA-directed DNA polymerase--properties and functions in oncogenic RNA viruses and cells. Prog Nucleic Acid Res Mol Biol. 1974;14(0):187–334. [PubMed] [Google Scholar]
  6. Humphries E. H., Temin H. M. Requirement for cell division for initiation of transcription of Rous sarcoma virus RNA. J Virol. 1974 Sep;14(3):531–546. doi: 10.1128/jvi.14.3.531-546.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Kacian D. L., Watson K. F., Burny A., Spiegelman S. Purification of the DNA polymerase of avian myeloblastosis virus. Biochim Biophys Acta. 1971 Sep 24;246(3):365–383. doi: 10.1016/0005-2787(71)90773-8. [DOI] [PubMed] [Google Scholar]
  8. Kang C. Y., Temin H. M. Reticuloendotheliosis virus nucleic acid sequences in cellular DNA. J Virol. 1974 Nov;14(5):1179–1188. doi: 10.1128/jvi.14.5.1179-1188.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [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. Mizutani S. DNA polymerases in RNA tumor viruses and possible "protoviruses". Am J Clin Pathol. 1973 Jul;60(1):19–24. doi: 10.1093/ajcp/60.1.19. [DOI] [PubMed] [Google Scholar]
  12. Mizutani S., Temin H. M. Lack of serological relationship among DNA polymerases of avian leukosis-sarcoma viruses, reticuloendotheliosis viruses, and chicken cells. J Virol. 1973 Sep;12(3):440–448. doi: 10.1128/jvi.12.3.440-448.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Mizutani S., Temin H. M. Specific serological relationships among partially purified DNA polymerases of avian leukosis-sarcoma viruses, reticuloendotheliosis viruses, and avian cells. J Virol. 1974 May;13(5):1020–1029. doi: 10.1128/jvi.13.5.1020-1029.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Moelling K. Characterization of reverse transcriptase and RNase H from friend-murine leukemia virus. Virology. 1974 Nov;62(1):46–59. doi: 10.1016/0042-6822(74)90302-x. [DOI] [PubMed] [Google Scholar]
  15. Moelling K., Gelderblom H., Pauli G., Friis R. A comparative study of the avian reticuloendotheliosis virus: relationship to murine leukemia virus and viruses of the avian sarcoma-leukosis complex. Virology. 1975 Jun;65(2):546–557. doi: 10.1016/0042-6822(75)90059-8. [DOI] [PubMed] [Google Scholar]
  16. Moelling K. Reverse transcriptase and RNase H: present in a murine virus and in both subunits of an avian virus. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 2):969–973. doi: 10.1101/sqb.1974.039.01.111. [DOI] [PubMed] [Google Scholar]
  17. Nakao T., Nakao M., Nagai F. Microdetermination of protein not affected by the presence of various buffers, sucrose, ATP, and eluates from polysaccharide derivatives. Anal Biochem. 1973 Oct;55(2):358–367. doi: 10.1016/0003-2697(73)90124-3. [DOI] [PubMed] [Google Scholar]
  18. Purchase H. G., Ludford C., Nazerian K., Cox H. W. A new group of oncogenic viruses: reticuloendotheliosis, chick syncytial, duck infectious anemia, and spleen necrosis viruses. J Natl Cancer Inst. 1973 Aug;51(2):489–499. [PubMed] [Google Scholar]
  19. Randerath K., Randerath E. Ion-exchange thin-layer chromatography. XIV. Separation of nucleotide sugars and nucleoside monophosphates on PEI-cellulose. Anal Biochem. 1965 Dec;13(3):575–579. doi: 10.1016/0003-2697(65)90356-8. [DOI] [PubMed] [Google Scholar]
  20. Temin H. M., Kassner V. K. Replication of reticuloendotheliosis viruses in cell culture: acute infection. J Virol. 1974 Feb;13(2):291–297. doi: 10.1128/jvi.13.2.291-297.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Temin H. M. On the origin of RNA tumor viruses. Annu Rev Genet. 1974;8:155–177. doi: 10.1146/annurev.ge.08.120174.001103. [DOI] [PubMed] [Google Scholar]
  22. Vogt V. M. Purification and further properties of single-strand-specific nuclease from Aspergillus oryzae. Eur J Biochem. 1973 Feb 15;33(1):192–200. doi: 10.1111/j.1432-1033.1973.tb02669.x. [DOI] [PubMed] [Google Scholar]

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

RESOURCES