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. 1990 Jun;64(6):2678–2686. doi: 10.1128/jvi.64.6.2678-2686.1990

A deletion in the Friend spleen focus-forming virus env gene is necessary for its product (gp55) to be leukemogenic.

N Watanabe 1, M Nishi 1, Y Ikawa 1, H Amanuma 1
PMCID: PMC249446  PMID: 2159537

Abstract

To determine the biological significance of the 585-base-pair deletion in the env gene of Friend spleen focus-forming virus (SFFV) encoding a leukemogenic glycoprotein (gp55), we examined the pathogenicity of a constructed mutant SFFV (SFFVDF). In the SFFVDF genome, the env deletion was filled in with the corresponding env sequence of Friend mink cell focus-forming virus, whereas the 6-base-pair duplication and the single base insertion near the 3' terminus of SFFV env remained intact. SFFVDF was nonpathogenic in adult mice. During passage of SFFVDF through newborn mice, we recovered various pathogenic variant SFFVs. Molecular analyses of variant SFFV genome DNAs revealed the presence of a distinct deletion in each env gene, which was similar but not identical to that in the wild-type SFFV env. Starting with the SFFVDF genome DNA, other mutant SFFV genome DNAs were constructed in which the sequence coding for the gp70/p15E proteolytic cleavage site present in the SFFVDF genome was modified by oligonucleotide-directed site-specific mutagenesis to prevent the cleavage. These mutant SFFVs were also nonpathogenic. These results indicate that for the pathogenic activity of gp55, a certain env deletion is necessary which causes production of a gp70-p15E fusion protein with an absence of at least the N-terminal one-third of the p15E-coding region.

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Selected References

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

  1. Adachi A., Sakai K., Kitamura N., Nakanishi S., Niwa O., Matsuyama M., Ishimoto A. Characterization of the env gene and long terminal repeat of molecularly cloned Friend mink cell focus-inducing virus DNA. J Virol. 1984 Jun;50(3):813–821. doi: 10.1128/jvi.50.3.813-821.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Amanuma H., Katori A., Obata M., Sagata N., Ikawa Y. Complete nucleotide sequence of the gene for the specific glycoprotein (gp55) of Friend spleen focus-forming virus. Proc Natl Acad Sci U S A. 1983 Jul;80(13):3913–3917. doi: 10.1073/pnas.80.13.3913. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Amanuma H., Watanabe N., Nishi M., Ikawa Y. Requirement of the single base insertion at the 3' end of the env-related gene of Friend spleen focus-forming virus for pathogenic activity and its effect on localization of the glycoprotein product (gp55). J Virol. 1989 Nov;63(11):4824–4833. doi: 10.1128/jvi.63.11.4824-4833.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bestwick R. K., Boswell B. A., Kabat D. Molecular cloning of biologically active Rauscher spleen focus-forming virus and the sequences of its env gene and long terminal repeat. J Virol. 1984 Sep;51(3):695–705. doi: 10.1128/jvi.51.3.695-705.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Clark S. P., Mak T. W. Complete nucleotide sequence of an infectious clone of Friend spleen focus-forming provirus: gp55 is an envelope fusion glycoprotein. Proc Natl Acad Sci U S A. 1983 Aug;80(16):5037–5041. doi: 10.1073/pnas.80.16.5037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gliniak B. C., Kabat D. Leukemogenic membrane glycoprotein encoded by Friend spleen focus-forming virus: transport to cell surfaces and shedding are controlled by disulfide-bonded dimerization and by cleavage of a hydrophobic membrane anchor. J Virol. 1989 Sep;63(9):3561–3568. doi: 10.1128/jvi.63.9.3561-3568.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hankins W. D., Troxler D. Polycythemia- and anemia-inducing erythroleukemia viruses exhibit differential erythroid transforming effects in vitro. Cell. 1980 Dec;22(3):693–699. doi: 10.1016/0092-8674(80)90545-0. [DOI] [PubMed] [Google Scholar]
  8. Kilpatrick D. R., Srinivas R. V., Stephens E. B., Compans R. W. Effects of deletion of the cytoplasmic domain upon surface expression and membrane stability of a viral envelope glycoprotein. J Biol Chem. 1987 Nov 25;262(33):16116–16121. [PubMed] [Google Scholar]
  9. Kunkel T. A., Roberts J. D., Zakour R. A. Rapid and efficient site-specific mutagenesis without phenotypic selection. Methods Enzymol. 1987;154:367–382. doi: 10.1016/0076-6879(87)54085-x. [DOI] [PubMed] [Google Scholar]
  10. Li J. P., Bestwick R. K., Spiro C., Kabat D. The membrane glycoprotein of Friend spleen focus-forming virus: evidence that the cell surface component is required for pathogenesis and that it binds to a receptor. J Virol. 1987 Sep;61(9):2782–2792. doi: 10.1128/jvi.61.9.2782-2792.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Linemeyer D. L., Menke J. G., Ruscetti S. K., Evans L. H., Scolnick E. M. Envelope gene sequences which encode the gp52 protein of spleen focus-forming virus are required for the induction of erythroid cell proliferation. J Virol. 1982 Jul;43(1):223–233. doi: 10.1128/jvi.43.1.223-233.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Lyles D. S., McConnell K. A. Subcellular localization of the env-related glycoproteins in Friend erythroleukemia cells. J Virol. 1981 Jul;39(1):263–272. doi: 10.1128/jvi.39.1.263-272.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Mager D. L., Mak T. W., Bernstein A. Quantitative colony method for tumorigenic cells transformed by two distinct strains of Friend leukemia virus. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1703–1707. doi: 10.1073/pnas.78.3.1703. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Obata M., Amanuma H., Harada Y., Sagata N., Ikawa Y. env-Related leukemogenic genes (gp55 genes) of two closely related polycythemic strains of Friend spleen focus-forming virus possess different recombination points with an endogenous mink cell focus-forming virus env gene. Virology. 1984 Jul 30;136(2):435–438. doi: 10.1016/0042-6822(84)90179-x. [DOI] [PubMed] [Google Scholar]
  15. Ostertag W., Stocking C., Johnson G. R., Kluge N., Kollek R., Franz T., Hess N. Transforming genes and target cells of murine spleen focus-forming viruses. Adv Cancer Res. 1987;48:193–355. doi: 10.1016/s0065-230x(08)60693-4. [DOI] [PubMed] [Google Scholar]
  16. Pinter A., Honnen W. J. Biochemical characterization of cell-associated and extracellular products of the Friend spleen focus-forming virus env gene. Virology. 1989 Nov;173(1):136–143. doi: 10.1016/0042-6822(89)90229-8. [DOI] [PubMed] [Google Scholar]
  17. Pinter A., Honnen W. J., Li J. S. Studies with inhibitors of oligosaccharide processing indicate a functional role for complex sugars in the transport and proteolysis of Friend mink cell focus-inducing murine leukemia virus envelope proteins. Virology. 1984 Jul 15;136(1):196–210. doi: 10.1016/0042-6822(84)90259-9. [DOI] [PubMed] [Google Scholar]
  18. Pinter A., Honnen W. J. O-linked glycosylation of retroviral envelope gene products. J Virol. 1988 Mar;62(3):1016–1021. doi: 10.1128/jvi.62.3.1016-1021.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Pinter A., Honnen W. J. The mature form of the Friend spleen focus-forming virus envelope protein, gp65, is efficiently secreted from cells. Virology. 1985 Jun;143(2):646–650. doi: 10.1016/0042-6822(85)90406-4. [DOI] [PubMed] [Google Scholar]
  20. Rowe W. P., Pugh W. E., Hartley J. W. Plaque assay techniques for murine leukemia viruses. Virology. 1970 Dec;42(4):1136–1139. doi: 10.1016/0042-6822(70)90362-4. [DOI] [PubMed] [Google Scholar]
  21. Ruscetti S., Wolff L. Spleen focus-forming virus: relationship of an altered envelope gene to the development of a rapid erythroleukemia. Curr Top Microbiol Immunol. 1984;112:21–44. doi: 10.1007/978-3-642-69677-0_2. [DOI] [PubMed] [Google Scholar]
  22. Ruta M., Bestwick R., Machida C., Kabat D. Loss of leukemogenicity caused by mutations in the membrane glycoprotein structural gene of Friend spleen focus-forming virus. Proc Natl Acad Sci U S A. 1983 Aug;80(15):4704–4708. doi: 10.1073/pnas.80.15.4704. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Ruta M., Kabat D. Plasma membrane glycoproteins encoded by cloned Rauscher and Friend spleen focus-forming viruses. J Virol. 1980 Sep;35(3):844–853. doi: 10.1128/jvi.35.3.844-853.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Srinivas R. V., Compans R. W. Glycosylation and intracellular transport of spleen focus-forming virus glycoproteins. Virology. 1983 Mar;125(2):274–286. doi: 10.1016/0042-6822(83)90201-5. [DOI] [PubMed] [Google Scholar]
  26. Wendling F., Moreau-Gachelin F., Tambourin P. Emergence of tumorigenic cells during the course of Friend virus leukemias. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3614–3618. doi: 10.1073/pnas.78.6.3614. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Wigler M., Pellicer A., Silverstein S., Axel R. Biochemical transfer of single-copy eucaryotic genes using total cellular DNA as donor. Cell. 1978 Jul;14(3):725–731. doi: 10.1016/0092-8674(78)90254-4. [DOI] [PubMed] [Google Scholar]
  28. Wolff L., Hubbert N., Ruscetti S. Structural analysis of the spleen focus-forming virus envelope gene product. Virology. 1984 Mar;133(2):376–385. doi: 10.1016/0042-6822(84)90403-3. [DOI] [PubMed] [Google Scholar]
  29. Wolff L., Kaminchik J., Hankins W. D., Ruscetti S. K. Sequence comparisons of the anemia- and polycythemia-inducing strains of Friend spleen focus-forming virus. J Virol. 1985 Feb;53(2):570–578. doi: 10.1128/jvi.53.2.570-578.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Wolff L., Scolnick E., Ruscetti S. Envelope gene of the Friend spleen focus-forming virus: deletion and insertions in 3' gp70/p15E-encoding region have resulted in unique features in the primary structure of its protein product. Proc Natl Acad Sci U S A. 1983 Aug;80(15):4718–4722. doi: 10.1073/pnas.80.15.4718. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Wolff L., Tambourin P., Ruscetti S. Induction of the autonomous stage of transformation in erythroid cells infected with SFFV: helper virus is not required. Virology. 1986 Jul 15;152(1):272–276. doi: 10.1016/0042-6822(86)90393-4. [DOI] [PubMed] [Google Scholar]

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