Summary
The DNA polymerase of the L-cell virion (LCV) was partially purified by chromatography on DEAE cellulose. This enzyme transcribed poly(A) · oligo (dT), poly(C) · oligo(dG), and poly(Cm) · oligo(dG), had a molecular weight of 77,000 daltons and reacted like other murine viral RNA directed DNA polymerases to anti reverse transcriptase specific IgG preparations indicating that it was probably a typical murine viral reverse transcriptase. In addition, like other partially purified mammalian viral reverse transcriptases the LCV DNA polymerase exhibited template independent primer stimulated DNA synthesis. The significance of these results to the unusual endogenous activity of the LCV is discussed.
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References
Abrell, J. W., Gallo, R. C.: Purification, characterisation and comparison of the DNA polymerases from two primate RNA tumour viruses. J. Virol.12, 431–439 (1973).
Abrell, J. W., Reitz, M. S., Gallo, R. C.: Transcription of 70s RNA by DNA Polymerases from mammalian RNA viruses. J. Virol.16, 1566–1574 (1975).
Ashley, R. L., Cardiff, R. D., Manning, J. S.: Characterisation of a terminal deoxynucleotidyl transferase activity in mouse mammary tumour virus. Virology77, 367–375 (1977).
Bhalla, R. B., Schwartz, M. K., Modak, M. J.: Selective inhibition of terminal deoxynucleotidyl transferase by ATP and its application in the detection of TdT in human leukemia. Biochem. biophys. Res. Commun.76, 1056–1061 (1977).
Bradford, M. M.: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem.72, 248–254 (1976).
Chang, L. M. S., Bollum, F. J.: Deoxynucleotide-polymerizing enzymes of calf thymus gland V homogenous terminal deoxynucleotidyl transferase. J. biol. Chem.246, 909–916 (1971).
Dales, S., Howatson, A. F.: Virus like particles in association with L stain cells. Cancer Res.21, 192–197 (1961).
Gerard, G. F., Rottman, F., Green, M.: Poly(2′-O-methylcytidylate) oligode-oxyguanylate as a template for the RNA directed DNA polymerase in RNA tumour virus particles and a specific probe for the RNA directed enzyme in transformed murine cells. Biochem.13, 1632–1641 (1974).
Green, M., Gerard, G. F.: RNA directed DNA polymerase-properties and functions in oncogenic RNA viruses and cells. Prog. Nucleic Acid. Res. Mol. Biol.14, 187–334 (1974).
Hallinan, F. M., Lee, S. H. S., Rozee, K. R.: Demonstration of an unusual DNA polymerase activity associated with the L cell virion. J. gen. Virol.42, 521–532 (1979).
Kacian, D. L.: Methods for assaying reverse transcriptase. Methods in Virol.6, 143–184 (1977).
Kindig, D. A., Kirsten, W. H.: Virus like particles in established murine cell lines; electron microscopic observations. Science155, 1543–1545 (1967).
Luftig, R. B., McMillan, P. N., Gudger, M.: Quantitation of endogenous C type virion production in several murine cell lines. J. Virol.14, 1017–1021 (1974).
Marcus, S. L., Sarkar, N. H.: Retroviral ‘Terminal deoxynucleotidyl transferase’ activity is reverse transcription. Virology84, 247–259 (1978).
Martin, R. G., Ames, B. N.: A method for determining the sedimentation behaviour of enzymes; application to protein mixtures. J. biol. Chem.236, 1372–1379 (1961).
Mizutani, S., Temin, H. M.: Purification and properties of spleen necrosis virus DNA polymerase. J. Virol.16, 797–806 (1975).
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.252, 11–19 (1977).
Modak, M. J.: Pyridoxal 5′ phosphate. A selective inhibitor of oncornaviral DNA polymerases. Biochem. biophys. Res. Commun.71, 180–187 (1976).
Nichols, J. L., Quade, K., Luftig, R. B.: Physicochemical studies on L cell virions. J. Virol.11, 432–440 (1973).
Nichols, J. L., Waddell, M., Comparison of free and 80s RNA associated RNA's of mouse L cell virions. Nature New. Biol.243, 236–238 (1973).
Pharmacia Manual (1977) Protein A, Protein A-Sepharose CL-4B.
Sarngadharan, M. G., Allaudeen, H. S., Gallo, R. C.: In:Busch, H. (ed.), Reverse transcriptase of RNA tumour viruses and animal cells in Methods in Cancer Res., Vol. 12, 3–47. New York: Academic Press 1976.
Srivastava, B. I. S., Minowada, J.: Terminal deoxynucleotidyl transferase activity and cell surface antigens of two unique cell lines (NALM-1 and BALM-2) of human leukemic origin. Int. J. Cancer20, 119–205 (1977).
Strickland, J. E., Hellman, A.: Detection of nanogram quantities of retroviral RNA by assay with reverse transcriptase. Anal. Biochem.82, 342–347 (1977).
Weber, G. H., Deeney, A. O'C., Beaudreau, G. S.: Isolation of DNA and DNA polymerase from MC29 tumour virus. Biochim. biophys. Acta299, 8–16 (1973).
Wu, A. M., Cetta, A.: On the stimulation of viral DNA polymerase activity by non ionic detergent. Biochem.14, 789–795 (1975).
Wu, A. M., Gallo, R. C.: Reverse transcriptase. CRC Critical Reviews in Biochem.3, 289–347 (1975).
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Hallinan, F.M., Lee, S.H.S. & Rozee, K.R. Isolation and characterisation of the L cell virion DNA polymerase. Archives of Virology 69, 71–83 (1981). https://doi.org/10.1007/BF01315267
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DOI: https://doi.org/10.1007/BF01315267