Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1996 Mar;70(3):1768–1774. doi: 10.1128/jvi.70.3.1768-1774.1996

Identification of a subdomain in the Moloney murine leukemia virus envelope protein involved in receptor binding.

A J MacKrell 1, N W Soong 1, C M Curtis 1, W F Anderson 1
PMCID: PMC190002  PMID: 8627699

Abstract

We have mutated amino acids within the receptor-binding domain of Moloney murine leukemia virus envelope in order to identify residues involved in receptor binding. Analysis of mutations in the region of amino acids 81 to 88 indicates that this region is important for specific envelope-receptor interactions. None of the aspartate 84 (D-84) mutants studied bind measurably, although they are efficiently incorporated into particles. D-84 mutants have titers that correspond to the severity of the substitution. This observation suggests that D-84 may provide a direct receptor contact. Mutations in the other charged amino acids in this domain (R-83, E-86, and E-87) yield titers similar to those of wild-type envelope, but the affinity of the mutant envelope in the binding assay is decreased by nonconservative substitutions in parallel to the severity of the change. These other amino acids may either provide secondary receptor contacts or assist in maintaining a structure in the domain that favors efficient binding. We also studied other regions of high hydrophilicity. Our initial characterization indicates that amino acids 106 to 111 and 170 to 188 do not play a major role in receptor binding. Measurements of relative binding affinity and titer indicate that most mutations in the region of amino acids 120 to 131 did not significantly affect receptor binding. However, SU encoded by mutants H123V, R124L, and C131A as well as C81A could not be detected in particles and therefore did not bind measurably. Therefore, the region encompassed by amino acids 81 to 88 appears to be directly involved in receptor binding.

Full Text

The Full Text of this article is available as a PDF (350.8 KB).

Selected References

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

  1. Albritton L. M., Kim J. W., Tseng L., Cunningham J. M. Envelope-binding domain in the cationic amino acid transporter determines the host range of ecotropic murine retroviruses. J Virol. 1993 Apr;67(4):2091–2096. doi: 10.1128/jvi.67.4.2091-2096.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Battini J. L., Danos O., Heard J. M. Receptor-binding domain of murine leukemia virus envelope glycoproteins. J Virol. 1995 Feb;69(2):713–719. doi: 10.1128/jvi.69.2.713-719.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Battini J. L., Heard J. M., Danos O. Receptor choice determinants in the envelope glycoproteins of amphotropic, xenotropic, and polytropic murine leukemia viruses. J Virol. 1992 Mar;66(3):1468–1475. doi: 10.1128/jvi.66.3.1468-1475.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Battini J. L., Kayman S. C., Pinter A., Heard J. M., Danos O. Role of N-linked glycosylation in the activity of the Friend murine leukemia virus SU protein receptor-binding domain. Virology. 1994 Jul;202(1):496–499. doi: 10.1006/viro.1994.1369. [DOI] [PubMed] [Google Scholar]
  5. Colicelli J., Lobel L. I., Goff S. P. A temperature-sensitive mutation constructed by "linker insertion" mutagenesis. Mol Gen Genet. 1985;199(3):537–539. doi: 10.1007/BF00330771. [DOI] [PubMed] [Google Scholar]
  6. Eiden M. V., Farrell K., Warsowe J., Mahan L. C., Wilson C. A. Characterization of a naturally occurring ecotropic receptor that does not facilitate entry of all ecotropic murine retroviruses. J Virol. 1993 Jul;67(7):4056–4061. doi: 10.1128/jvi.67.7.4056-4061.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Evans L. H., Morrison R. P., Malik F. G., Portis J., Britt W. J. A neutralizable epitope common to the envelope glycoproteins of ecotropic, polytropic, xenotropic, and amphotropic murine leukemia viruses. J Virol. 1990 Dec;64(12):6176–6183. doi: 10.1128/jvi.64.12.6176-6183.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Felkner R. H., Roth M. J. Mutational analysis of the N-linked glycosylation sites of the SU envelope protein of Moloney murine leukemia virus. J Virol. 1992 Jul;66(7):4258–4264. doi: 10.1128/jvi.66.7.4258-4264.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Graham F. L., van der Eb A. J. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology. 1973 Apr;52(2):456–467. doi: 10.1016/0042-6822(73)90341-3. [DOI] [PubMed] [Google Scholar]
  10. Gray K. D., Roth M. J. Mutational analysis of the envelope gene of Moloney murine leukemia virus. J Virol. 1993 Jun;67(6):3489–3496. doi: 10.1128/jvi.67.6.3489-3496.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Heard J. M., Danos O. An amino-terminal fragment of the Friend murine leukemia virus envelope glycoprotein binds the ecotropic receptor. J Virol. 1991 Aug;65(8):4026–4032. doi: 10.1128/jvi.65.8.4026-4032.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hopp T. P., Woods K. R. Prediction of protein antigenic determinants from amino acid sequences. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3824–3828. doi: 10.1073/pnas.78.6.3824. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hunter E., Swanstrom R. Retrovirus envelope glycoproteins. Curr Top Microbiol Immunol. 1990;157:187–253. doi: 10.1007/978-3-642-75218-6_7. [DOI] [PubMed] [Google Scholar]
  14. Kadan M. J., Sturm S., Anderson W. F., Eglitis M. A. Detection of receptor-specific murine leukemia virus binding to cells by immunofluorescence analysis. J Virol. 1992 Apr;66(4):2281–2287. doi: 10.1128/jvi.66.4.2281-2287.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kayman S. C., Kopelman R., Projan S., Kinney D. M., Pinter A. Mutational analysis of N-linked glycosylation sites of Friend murine leukemia virus envelope protein. J Virol. 1991 Oct;65(10):5323–5332. doi: 10.1128/jvi.65.10.5323-5332.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Linder M., Linder D., Hahnen J., Schott H. H., Stirm S. Localization of the intrachain disulfide bonds of the envelope glycoprotein 71 from Friend murine leukemia virus. Eur J Biochem. 1992 Jan 15;203(1-2):65–73. doi: 10.1111/j.1432-1033.1992.tb19828.x. [DOI] [PubMed] [Google Scholar]
  17. Linder M., Wenzel V., Linder D., Stirm S. Structural elements in glycoprotein 70 from polytropic Friend mink cell focus-inducing virus and glycoprotein 71 from ecotropic Friend murine leukemia virus, as defined by disulfide-bonding pattern and limited proteolysis. J Virol. 1994 Aug;68(8):5133–5141. doi: 10.1128/jvi.68.8.5133-5141.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Miller A. D., Rosman G. J. Improved retroviral vectors for gene transfer and expression. Biotechniques. 1989 Oct;7(9):980-2, 984-6, 989-90. [PMC free article] [PubMed] [Google Scholar]
  19. Morgan R. A., Nussbaum O., Muenchau D. D., Shu L., Couture L., Anderson W. F. Analysis of the functional and host range-determining regions of the murine ectropic and amphotropic retrovirus envelope proteins. J Virol. 1993 Aug;67(8):4712–4721. doi: 10.1128/jvi.67.8.4712-4721.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Nakamaye K. L., Eckstein F. Inhibition of restriction endonuclease Nci I cleavage by phosphorothioate groups and its application to oligonucleotide-directed mutagenesis. Nucleic Acids Res. 1986 Dec 22;14(24):9679–9698. doi: 10.1093/nar/14.24.9679. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Ott D., Friedrich R., Rein A. Sequence analysis of amphotropic and 10A1 murine leukemia viruses: close relationship to mink cell focus-inducing viruses. J Virol. 1990 Feb;64(2):757–766. doi: 10.1128/jvi.64.2.757-766.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Ott D., Rein A. Basis for receptor specificity of nonecotropic murine leukemia virus surface glycoprotein gp70SU. J Virol. 1992 Aug;66(8):4632–4638. doi: 10.1128/jvi.66.8.4632-4638.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Rein A., Schultz A. Different recombinant murine leukemia viruses use different cell surface receptors. Virology. 1984 Jul 15;136(1):144–152. doi: 10.1016/0042-6822(84)90255-1. [DOI] [PubMed] [Google Scholar]
  24. Russel M., Kidd S., Kelley M. R. An improved filamentous helper phage for generating single-stranded plasmid DNA. Gene. 1986;45(3):333–338. doi: 10.1016/0378-1119(86)90032-6. [DOI] [PubMed] [Google Scholar]
  25. Skov H., Andersen K. B. Mutational analysis of Moloney murine leukaemia virus surface protein gp70. J Gen Virol. 1993 Apr;74(Pt 4):707–714. doi: 10.1099/0022-1317-74-4-707. [DOI] [PubMed] [Google Scholar]
  26. Vogt M., Haggblom C., Swift S., Haas M. Specific sequences of the env gene determine the host range of two XC-negative viruses of the Rauscher virus complex. Virology. 1986 Oct 30;154(2):420–424. doi: 10.1016/0042-6822(86)90470-8. [DOI] [PubMed] [Google Scholar]
  27. Yoshimoto T., Yoshimoto E., Meruelo D. Identification of amino acid residues critical for infection with ecotropic murine leukemia retrovirus. J Virol. 1993 Mar;67(3):1310–1314. doi: 10.1128/jvi.67.3.1310-1314.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Yu H., Soong N., Anderson W. F. Binding kinetics of ecotropic (Moloney) murine leukemia retrovirus with NIH 3T3 cells. J Virol. 1995 Oct;69(10):6557–6562. doi: 10.1128/jvi.69.10.6557-6562.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES