Abstract
Genetic evidence indicates that the oncogenic potential of Rous sarcoma virus (RSV) resides in a single viral gene denoted src1,2. The product of this gene is a 60,000-molecular weight (Mr) phosphoprotein (pp60src)3–6 which is linked to the plasma membrane of RSV-infected cells7–9 by an interaction at its amino terminus10. This protein is associated with a protein kinase activity5,11 capable of phosphorylating tyrosine residues in acceptor proteins12–15. A closely related protein occurs in uninfected cells from a wide variety of vertebrates16–18 and is presumably encoded by a cellular gene that is alleged to be the progenitor of src16–19. Because detailed enzymological characterization of pp60src has been hampered by a lack of sufficient quantities of purified material, we have produced a bacterial plasmid capable of expressing in Escherichia coli a protein closely related to pp60src. We report here the construction of this vector, which directs the synthesis in E. coli of large quantities of a protein having the size and immunological properties characteristic of pp60src. In addition, we have demonstrated tyrosine-specific protein kinase activity associated with the bacterially derived protein, an observation which greatly strengthens the claim that the kinase activity previously observed to be associated with pp60src represents a bona fide property of the protein.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 51 print issues and online access
$199.00 per year
only $3.90 per issue
Buy this article
- Purchase on SpringerLink
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Hanafusa, H. in Comprehensive Virology Vol. 10, 401–483 (Plenum, New York, 1977).
Vogt, P. K. in Comprehensive Virology Vol. 10, 341–455 (Plenum, New York, 1977).
Brugge, J. S. & Erikson, R. L. Nature 269, 346–348 (1977).
Purchio, A. F., Erikson, E., Brugge, J. S. & Erikson, R. L. Proc. natn. Acad. Sci. U.S.A. 75, 1567–1571 (1978).
Levinson, A. D., Oppermann, H. O., Levintow, L., Varmus, H. E. & Bishop, J. M. Cell 15, 561–572 (1978).
Collett, M. S. et al. Proc. natn. Acad. Sci. U.S.A. 76, 3159–3163 (1979).
Willingham, M. C., Jay, G. & Pastan, I. Cell 18, 125–134 (1979).
Kruger, J. G., Wang, E. & Goldberg, A. R. Virology 101, 25–49 (1980).
Courtneidge, S. A., Levinson, A. D. & Bishop, J. M. Proc. natn. Acad. Sci. U.S.A. 77, 3783–3787 (1980).
Levinson, A. D., Courtneidge, S. A. & Bishop, J. M. Proc. natn. Acad. Sci. U.S.A. 78, 1624–1628 (1981).
Collett, M. S. & Erikson, R. L. Proc. natn. Acad. Sci. U.S.A. 75, 2021–2024 (1978).
Hunter, T. & Sefton, B. M. Proc. natn. Acad. Sci. U.S.A. 77, 1311–1315 (1980).
Sefton, B. M., Hunter, T., Beemon, K. & Eckhart, W. Cell 20, 807–816 (1980).
Collett, M., Purchio, A. F. & Erikson, R. L. Nature 285, 167–169 (1980).
Levinson, A. D., Oppermann, H. O., Varmus, H. E. & Bishop, J. M. J. biol. Chem. 255, 11973–11980 (1980).
Collett, M. S., Brugge, J. S. & Erikson, R. L. Cell 15, 1363–1369 (1978).
Oppermann, H. O., Levinson, A. D., Varmus, H. E., Levintow, L. & Bishop, J. M. Proc. natn. Acad. Sci. U.S.A. 76, 1804–1808 (1979).
Karess, R. E., Hayward, W. S. & Hanafusa, H. Proc. natn. Acad. Sci. U.S.A. 76, 3154–3158 (1979).
Spector, D., Varmus, H. E. & Bishop, M. J. Proc. natn. Acad. Sci. U.S.A. 75, 4102–4106 (1978).
Goeddel, D. G. et al. Nature 281, 544–548 (1979).
De Boer, H. A. et al. in Promoters, Structure and Function (eds Chamberlain, M. J. & Rodriguez, R.) (Praeger Scientific, 1981).
Czernilofsky, A. P. et al. Nature 287, 198–203 (1980).
Yanofsky, C. Nature 289, 751–758 (1981).
Cleveland, D. W. et al. J. biol. Chem. 252, 1102–1106 (1977).
Erikson, R. L., Collett, M. S., Erikson, E. & Purchio, A. F. Proc. natn. Acad. Sci. U.S.A. 76, 6260–6264 (1979).
Rubin, C. S. & Rosen, O. M. A. Rev. Biochem. 44, 831–887 (1975).
Rahmsdorf, H. J. et al. Proc. natn. Acad. Sci. U.S.A. 71, 586–589 (1974).
Khandelwal, R. L., Spearman, T. N. & Hamilton, I. R. FEBS Lett. 31, 246–252 (1973).
Wang, J. & Koshland, D. E. J. biol. Chem. 253, 7605–7608 (1978).
Garnak, M. & Reeves, H. C. J. biol. Chem. 254, 7915–7920 (1979).
Wang, J. & Koshland, D. E. J. biol. Chem. 256, 4640–4648 (1981).
Smith, H. Meth. Enzym. 65, 371–380 (1980).
Backman, K., Ptashne, M. & Gilbert, W. Proc. natn. Acad. Sci. U.S.A. 73, 4174–4178 (1976).
Laemmli, U. K. Nature 227, 680 (1970).
Gilmer, T. M. & Erikson, R. L. Nature 294, 771–773 (1981).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
McGrath, J., Levinson, A. Bacterial expression of an enzymatically active protein encoded by RSV src gene. Nature 295, 423–425 (1982). https://doi.org/10.1038/295423a0
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1038/295423a0
Comments
By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.