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Journal of Virology logoLink to Journal of Virology
. 1981 Jan;37(1):274–283. doi: 10.1128/jvi.37.1.274-283.1981

Endodeoxyribonuclease activity associated with Rauscher murine leukemia virus.

J J Kopchick, J Harless, B S Geisser, R Killam, R R Hewitt, R B Arlinghaus
PMCID: PMC171005  PMID: 6260982

Abstract

Preparations of purified Rauscher murine leukemia virus were found to contain an endodeoxyribonuclease after disruption of the virus with nonionic detergents. The enzyme makes single-strand breaks in linear or covalently closed circular phage double-stranded DNA molecules. The enzyme was partially purified by ion-exchange chromatography on DEAE- and carboxymethyl-Sepharose columns followed by electrophoresis in DNA-containing polyacrylamide gels. The enzyme was separated from reverse transcriptase (p80pol), and the final endonuclease preparation contained no detectable reverse transcriptase activity. The DEAE-Sepharose column-purified endonuclease activity contained a polypeptide of about 40,000 Mr that we term p40. Peptide mapping experiments demonstrated that p40 shares methionine-labeled tryptic peptides with Pr200gag-pol and Pr135pol. Six major methionine-labeled tryptic peptides derived from p40 were found in Pr200gag-pol, but only five of these were detected in Pr135pol. The four core proteins (p30, p15, pp12, and p10) and p80pol plus p40 account for most, but not all, of the peptide sequences of Pr200gag-pol. The endonuclease-associated p40 is similar in size and precursor origin to the avian retrovirus-coded endonuclease (p32). In view of these similarities to the avian p32 endonuclease and its association with partially purified Rauscher murine leukemia virus-associated endonuclease preparations, we propose that p40 is the Rauscher murine leukemia virus-coded endonuclease.

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Selected References

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  1. Arcement L. J., Karshin W. L., Naso R. B., Arlinghaus R. B. "gag" polyprotein precursors of Rauscher murine leukemia virus. Virology. 1977 Sep;81(2):284–297. doi: 10.1016/0042-6822(77)90145-3. [DOI] [PubMed] [Google Scholar]
  2. Grandgenett D. P., Golomb M., Vora A. C. Activation of an Mg2+-dependent DNA endonuclease of avian myeloblastosis virus alpha beta DNA polymerase by in vitro proteolytic cleavage. J Virol. 1980 Jan;33(1):264–271. doi: 10.1128/jvi.33.1.264-271.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Grandgenett D. P., Vora A. C., Schiff R. D. A 32,000-dalton nucleic acid-binding protein from avian retravirus cores possesses DNA endonuclease activity. Virology. 1978 Aug;89(1):119–132. doi: 10.1016/0042-6822(78)90046-6. [DOI] [PubMed] [Google Scholar]
  4. Gray H. B., Jr, Ostrander D. A., Hodnett J. L., Legerski R. J., Robberson D. L. Extracellular nucleases of Pseudomonas BAL 31. I. Characterization of single strand-specific deoxyriboendonuclease and double-strand deoxyriboexonuclease activities. Nucleic Acids Res. 1975 Sep;2(9):1459–1492. doi: 10.1093/nar/2.9.1459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Grdina D. J., Lohman P. H., Hewitt R. R. A fluorometric method for the detection of endodeoxyribonuclease on DNA-polyacrylamide gels. Anal Biochem. 1973 Jan;51(1):255–264. doi: 10.1016/0003-2697(73)90473-9. [DOI] [PubMed] [Google Scholar]
  6. Hewitt R., Suit J. C., Billen D. Utilization of 5-bromouracil by thymineless bacteria. J Bacteriol. 1967 Jan;93(1):86–89. doi: 10.1128/jb.93.1.86-89.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Jamjoom G. A., Naso R. B., Arlinghaus R. B. Further characterization of intracellular precursor polyproteins of Rauscher leukemia virus. Virology. 1977 May 1;78(1):11–34. doi: 10.1016/0042-6822(77)90075-7. [DOI] [PubMed] [Google Scholar]
  8. Jamjoom G. A., Naso R. B., Arlinghaus R. B. Selective decrease in the rate of cleavage of an intracellular precursor to Rauscher leukemia virus p30 by treatment of infected cells with actinomycin D. J Virol. 1976 Sep;19(3):1054–1072. doi: 10.1128/jvi.19.3.1054-1072.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Karshin W. L., Arcement L. J., Naso R. B., Arlinghaus R. B. Common precursor for Rauscher leukemia virus gp69/71, p15(E), and p12(E). J Virol. 1977 Sep;23(3):787–798. doi: 10.1128/jvi.23.3.787-798.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kopchick J. J., Jamjoom G. A., Watson K. F., Arlinghaus R. B. Biosynthesis of reverse transcriptase from Rauscher murine leukemia virus by synthesis and cleavage of a gag-pol read-through viral precursor polyprotein. Proc Natl Acad Sci U S A. 1978 Apr;75(4):2016–2020. doi: 10.1073/pnas.75.4.2016. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kopchick J. J., Karshin W. L., Arlinghaus R. B. Tryptic peptide analysis of gag and gag-pol gene products of Rauscher murine leukemia virus. J Virol. 1979 May;30(2):610–623. doi: 10.1128/jvi.30.2.610-623.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Love J. D., Hewitt R. R. The relationship between human serum and human pancreatic DNase I. J Biol Chem. 1979 Dec 25;254(24):12588–12594. [PubMed] [Google Scholar]
  14. McDonell M. W., Simon M. N., Studier F. W. Analysis of restriction fragments of T7 DNA and determination of molecular weights by electrophoresis in neutral and alkaline gels. J Mol Biol. 1977 Feb 15;110(1):119–146. doi: 10.1016/s0022-2836(77)80102-2. [DOI] [PubMed] [Google Scholar]
  15. Mizutani S., Boettiger D., Temin H. M. A DNA-depenent DNA polymerase and a DNA endonuclease in virions of Rous sarcoma virus. Nature. 1970 Oct 31;228(5270):424–427. doi: 10.1038/228424a0. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Murphy E. C., Jr, Arlinghaus R. B. Effect of canavanine on murine retrovirus polypeptide formation. J Virol. 1980 Mar;33(3):954–961. doi: 10.1128/jvi.33.3.954-961.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Murphy E. C., Jr, Kopchick J. J., Watson K. F., Arlinghaus R. B. Cell-free synthesis of a precursor polyprotein containing both gag and pol gene products by Rauscher murine leukemia virus 35S RNA. Cell. 1978 Feb;13(2):359–369. doi: 10.1016/0092-8674(78)90204-0. [DOI] [PubMed] [Google Scholar]
  19. Naso R. B., Arcement L. J., Arlinghaus R. B. Biosynthesis of Rauscher leukemia viral proteins. Cell. 1975 Jan;4(1):31–36. doi: 10.1016/0092-8674(75)90130-0. [DOI] [PubMed] [Google Scholar]
  20. Naso R. B., Karshin W. L., Wu Y. H., Arlinghaus R. B. Characterization of 40,000- and 25,000-dalton intermediate precursors to Rauscher murine leukemia virus gag gene products. J Virol. 1979 Oct;32(1):187–198. doi: 10.1128/jvi.32.1.187-198.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Oppermann H., Bishop J. M., Varmus H. E., Levintow L. A joint produce of the genes gag and pol of avian sarcoma virus: a possible precursor of reverse transcriptase. Cell. 1977 Dec;12(4):993–1005. doi: 10.1016/0092-8674(77)90164-7. [DOI] [PubMed] [Google Scholar]
  22. Paterson B. M., Marciani D. J., Papas T. S. Cell-free synthesis of the precursor polypeptide for avian myeloblastosis virus DNA polymerase. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4951–4954. doi: 10.1073/pnas.74.11.4951. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Schiff R. D., Grandgenett D. P. Virus-coded origin of a 32,000-dalton protein from avian retrovirus cores: structural relatedness of p32 and the beta polypeptide of the avian retrovirus DNA polymerase. J Virol. 1978 Oct;28(1):279–291. doi: 10.1128/jvi.28.1.279-291.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Syrewicz J. J., Naso R. B., Wang C. S., Arlinghaus R. B. Purification of large amounts of murine ribonucleic acid tumor viruses produced in roller bottle cultures. Appl Microbiol. 1972 Sep;24(3):488–494. doi: 10.1128/am.24.3.488-494.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. 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]

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