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. 1997 Jan;71(1):766–770. doi: 10.1128/jvi.71.1.766-770.1997

Identification of a cytoplasmic Tyr-X-X-Leu motif essential for down regulation of the human cell receptor CD46 in persistent measles virus infection.

S Yant 1, A Hirano 1, T C Wong 1
PMCID: PMC191115  PMID: 8985414

Abstract

To investigate the sequence requirements for measles virus (MV)-induced receptor down regulation, we transfected the human CD46 gene into simian cells persistently infected by the Biken strain of MV. Surface expression of CD46 is drastically reduced in these cells. Deletion analysis has shown that the juxtamembrane region of the CD46 cytoplasmic domain is essential for down regulation. Deleting a Tyr-Arg-Tyr-Leu sequence in this region or changing these residues to Ala prevents CD46 down regulation from the infected cell surface. Alanine-scanning mutagenesis has identified two amino acid residues, Tyr and Leu, forming a Tyr-X-X-Leu motif critical for CD46 down regulation. Mutations that prevent CD46 down regulation enhance syncytium formation. These results indicate that CD46 down regulation limits the cytopathic effects in a persistent MV infection and that CD46 down regulation requires a cytoplasmic Tyr-X-X-Leu sequence which resembles known motifs for membrane protein trafficking and receptor signalling.

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Selected References

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  1. Chen W. S., Lazar C. S., Lund K. A., Welsh J. B., Chang C. P., Walton G. M., Der C. J., Wiley H. S., Gill G. N., Rosenfeld M. G. Functional independence of the epidermal growth factor receptor from a domain required for ligand-induced internalization and calcium regulation. Cell. 1989 Oct 6;59(1):33–43. doi: 10.1016/0092-8674(89)90867-2. [DOI] [PubMed] [Google Scholar]
  2. Crise B., Buonocore L., Rose J. K. CD4 is retained in the endoplasmic reticulum by the human immunodeficiency virus type 1 glycoprotein precursor. J Virol. 1990 Nov;64(11):5585–5593. doi: 10.1128/jvi.64.11.5585-5593.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Dörig R. E., Marcil A., Chopra A., Richardson C. D. The human CD46 molecule is a receptor for measles virus (Edmonston strain). Cell. 1993 Oct 22;75(2):295–305. doi: 10.1016/0092-8674(93)80071-l. [DOI] [PubMed] [Google Scholar]
  4. Flaswinkel H., Barner M., Reth M. The tyrosine activation motif as a target of protein tyrosine kinases and SH2 domains. Semin Immunol. 1995 Feb;7(1):21–27. doi: 10.1016/1044-5323(95)90004-7. [DOI] [PubMed] [Google Scholar]
  5. Futter C. E., Connolly C. N., Cutler D. F., Hopkins C. R. Newly synthesized transferrin receptors can be detected in the endosome before they appear on the cell surface. J Biol Chem. 1995 May 5;270(18):10999–11003. doi: 10.1074/jbc.270.18.10999. [DOI] [PubMed] [Google Scholar]
  6. Gellin B. G., Katz S. L. Measles: state of the art and future directions. J Infect Dis. 1994 Nov;170 (Suppl 1):S3–14. doi: 10.1093/infdis/170.supplement_1.s3. [DOI] [PubMed] [Google Scholar]
  7. Gerlier D., Loveland B., Varior-Krishnan G., Thorley B., McKenzie I. F., Rabourdin-Combe C. Measles virus receptor properties are shared by several CD46 isoforms differing in extracellular regions and cytoplasmic tails. J Gen Virol. 1994 Sep;75(Pt 9):2163–2171. doi: 10.1099/0022-1317-75-9-2163. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Hirano A. Subacute sclerosing panencephalitis virus dominantly interferes with replication of wild-type measles virus in a mixed infection: implication for viral persistence. J Virol. 1992 Apr;66(4):1891–1898. doi: 10.1128/jvi.66.4.1891-1898.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hirano A., Yant S., Iwata K., Korte-Sarfaty J., Seya T., Nagasawa S., Wong T. C. Human cell receptor CD46 is down regulated through recognition of a membrane-proximal region of the cytoplasmic domain in persistent measles virus infection. J Virol. 1996 Oct;70(10):6929–6936. doi: 10.1128/jvi.70.10.6929-6936.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Ho S. N., Hunt H. D., Horton R. M., Pullen J. K., Pease L. R. Site-directed mutagenesis by overlap extension using the polymerase chain reaction. Gene. 1989 Apr 15;77(1):51–59. doi: 10.1016/0378-1119(89)90358-2. [DOI] [PubMed] [Google Scholar]
  12. Höning S., Hunziker W. Cytoplasmic determinants involved in direct lysosomal sorting, endocytosis, and basolateral targeting of rat lgp120 (lamp-I) in MDCK cells. J Cell Biol. 1995 Feb;128(3):321–332. doi: 10.1083/jcb.128.3.321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Karp C. L., Wysocka M., Wahl L. M., Ahearn J. M., Cuomo P. J., Sherry B., Trinchieri G., Griffin D. E. Mechanism of suppression of cell-mediated immunity by measles virus. Science. 1996 Jul 12;273(5272):228–231. doi: 10.1126/science.273.5272.228. [DOI] [PubMed] [Google Scholar]
  14. Krantic S., Gimenez C., Rabourdin-Combe C. Cell-to-cell contact via measles virus haemagglutinin-CD46 interaction triggers CD46 downregulation. J Gen Virol. 1995 Nov;76(Pt 11):2793–2800. doi: 10.1099/0022-1317-76-11-2793. [DOI] [PubMed] [Google Scholar]
  15. Lecouturier V., Fayolle J., Caballero M., Carabaña J., Celma M. L., Fernandez-Muñoz R., Wild T. F., Buckland R. Identification of two amino acids in the hemagglutinin glycoprotein of measles virus (MV) that govern hemadsorption, HeLa cell fusion, and CD46 downregulation: phenotypic markers that differentiate vaccine and wild-type MV strains. J Virol. 1996 Jul;70(7):4200–4204. doi: 10.1128/jvi.70.7.4200-4204.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Liszewski M. K., Post T. W., Atkinson J. P. Membrane cofactor protein (MCP or CD46): newest member of the regulators of complement activation gene cluster. Annu Rev Immunol. 1991;9:431–455. doi: 10.1146/annurev.iy.09.040191.002243. [DOI] [PubMed] [Google Scholar]
  17. Lublin D. M., Liszewski M. K., Post T. W., Arce M. A., Le Beau M. M., Rebentisch M. B., Lemons L. S., Seya T., Atkinson J. P. Molecular cloning and chromosomal localization of human membrane cofactor protein (MCP). Evidence for inclusion in the multigene family of complement-regulatory proteins. J Exp Med. 1988 Jul 1;168(1):181–194. doi: 10.1084/jem.168.1.181. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Manchester M., Liszewski M. K., Atkinson J. P., Oldstone M. B. Multiple isoforms of CD46 (membrane cofactor protein) serve as receptors for measles virus. Proc Natl Acad Sci U S A. 1994 Mar 15;91(6):2161–2165. doi: 10.1073/pnas.91.6.2161. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Manchester M., Valsamakis A., Kaufman R., Liszewski M. K., Alvarez J., Atkinson J. P., Lublin D. M., Oldstone M. B. Measles virus and C3 binding sites are distinct on membrane cofactor protein (CD46). Proc Natl Acad Sci U S A. 1995 Mar 14;92(6):2303–2307. doi: 10.1073/pnas.92.6.2303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Monlauzeur L., Rajasekaran A., Chao M., Rodriguez-Boulan E., Le Bivic A. A cytoplasmic tyrosine is essential for the basolateral localization of mutants of the human nerve growth factor receptor in Madin-Darby canine kidney cells. J Biol Chem. 1995 May 19;270(20):12219–12225. doi: 10.1074/jbc.270.20.12219. [DOI] [PubMed] [Google Scholar]
  21. Moscona A., Peluso R. W. Fusion properties of cells persistently infected with human parainfluenza virus type 3: participation of hemagglutinin-neuraminidase in membrane fusion. J Virol. 1991 Jun;65(6):2773–2777. doi: 10.1128/jvi.65.6.2773-2777.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Naniche D., Varior-Krishnan G., Cervoni F., Wild T. F., Rossi B., Rabourdin-Combe C., Gerlier D. Human membrane cofactor protein (CD46) acts as a cellular receptor for measles virus. J Virol. 1993 Oct;67(10):6025–6032. doi: 10.1128/jvi.67.10.6025-6032.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Naniche D., Wild T. F., Rabourdin-Combe C., Gerlier D. Measles virus haemagglutinin induces down-regulation of gp57/67, a molecule involved in virus binding. J Gen Virol. 1993 Jun;74(Pt 6):1073–1079. doi: 10.1099/0022-1317-74-6-1073. [DOI] [PubMed] [Google Scholar]
  24. Ohno H., Stewart J., Fournier M. C., Bosshart H., Rhee I., Miyatake S., Saito T., Gallusser A., Kirchhausen T., Bonifacino J. S. Interaction of tyrosine-based sorting signals with clathrin-associated proteins. Science. 1995 Sep 29;269(5232):1872–1875. doi: 10.1126/science.7569928. [DOI] [PubMed] [Google Scholar]
  25. Peters C., Braun M., Weber B., Wendland M., Schmidt B., Pohlmann R., Waheed A., von Figura K. Targeting of a lysosomal membrane protein: a tyrosine-containing endocytosis signal in the cytoplasmic tail of lysosomal acid phosphatase is necessary and sufficient for targeting to lysosomes. EMBO J. 1990 Nov;9(11):3497–3506. doi: 10.1002/j.1460-2075.1990.tb07558.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Post T. W., Liszewski M. K., Adams E. M., Tedja I., Miller E. A., Atkinson J. P. Membrane cofactor protein of the complement system: alternative splicing of serine/threonine/proline-rich exons and cytoplasmic tails produces multiple isoforms that correlate with protein phenotype. J Exp Med. 1991 Jul 1;174(1):93–102. doi: 10.1084/jem.174.1.93. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Purcell D. F., Russell S. M., Deacon N. J., Brown M. A., Hooker D. J., McKenzie I. F. Alternatively spliced RNAs encode several isoforms of CD46 (MCP), a regulator of complement activation. Immunogenetics. 1991;33(5-6):335–344. doi: 10.1007/BF00216692. [DOI] [PubMed] [Google Scholar]
  28. Reithmeier R. A. Characterization and modeling of membrane proteins using sequence analysis. Curr Opin Struct Biol. 1995 Aug;5(4):491–500. doi: 10.1016/0959-440x(95)80034-4. [DOI] [PubMed] [Google Scholar]
  29. Schneider-Schaulies J., Dunster L. M., Kobune F., Rima B., ter Meulen V. Differential downregulation of CD46 by measles virus strains. J Virol. 1995 Nov;69(11):7257–7259. doi: 10.1128/jvi.69.11.7257-7259.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Schneider-Schaulies J., Schnorr J. J., Brinckmann U., Dunster L. M., Baczko K., Liebert U. G., Schneider-Schaulies S., ter Meulen V. Receptor usage and differential downregulation of CD46 by measles virus wild-type and vaccine strains. Proc Natl Acad Sci U S A. 1995 Apr 25;92(9):3943–3947. doi: 10.1073/pnas.92.9.3943. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Schneider-Schaulies J., Schnorr J. J., Schlender J., Dunster L. M., Schneider-Schaulies S., ter Meulen V. Receptor (CD46) modulation and complement-mediated lysis of uninfected cells after contact with measles virus-infected cells. J Virol. 1996 Jan;70(1):255–263. doi: 10.1128/jvi.70.1.255-263.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Schnorr J. J., Dunster L. M., Nanan R., Schneider-Schaulies J., Schneider-Schaulies S., ter Meulen V. Measles virus-induced down-regulation of CD46 is associated with enhanced sensitivity to complement-mediated lysis of infected cells. Eur J Immunol. 1995 Apr;25(4):976–984. doi: 10.1002/eji.1830250418. [DOI] [PubMed] [Google Scholar]
  33. Schäfer W., Stroh A., Berghöfer S., Seiler J., Vey M., Kruse M. L., Kern H. F., Klenk H. D., Garten W. Two independent targeting signals in the cytoplasmic domain determine trans-Golgi network localization and endosomal trafficking of the proprotein convertase furin. EMBO J. 1995 Jun 1;14(11):2424–2435. doi: 10.1002/j.1460-2075.1995.tb07240.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Seya T., Hara T., Matsumoto M., Akedo H. Quantitative analysis of membrane cofactor protein (MCP) of complement. High expression of MCP on human leukemia cell lines, which is down-regulated during cell differentiation. J Immunol. 1990 Jul 1;145(1):238–245. [PubMed] [Google Scholar]
  35. Shaw A. S., Gauen L. K., Zhu Y. Interactions of TCR tyrosine based activation motifs with tyrosine kinases. Semin Immunol. 1995 Feb;7(1):13–20. doi: 10.1016/1044-5323(95)90003-9. [DOI] [PubMed] [Google Scholar]
  36. Takebe Y., Seiki M., Fujisawa J., Hoy P., Yokota K., Arai K., Yoshida M., Arai N. SR alpha promoter: an efficient and versatile mammalian cDNA expression system composed of the simian virus 40 early promoter and the R-U5 segment of human T-cell leukemia virus type 1 long terminal repeat. Mol Cell Biol. 1988 Jan;8(1):466–472. doi: 10.1128/mcb.8.1.466. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Taussig R., Gilman A. G. Mammalian membrane-bound adenylyl cyclases. J Biol Chem. 1995 Jan 6;270(1):1–4. doi: 10.1074/jbc.270.1.1. [DOI] [PubMed] [Google Scholar]
  38. Voorhees P., Deignan E., van Donselaar E., Humphrey J., Marks M. S., Peters P. J., Bonifacino J. S. An acidic sequence within the cytoplasmic domain of furin functions as a determinant of trans-Golgi network localization and internalization from the cell surface. EMBO J. 1995 Oct 16;14(20):4961–4975. doi: 10.1002/j.1460-2075.1995.tb00179.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Wild T. F., Malvoisin E., Buckland R. Measles virus: both the haemagglutinin and fusion glycoproteins are required for fusion. J Gen Virol. 1991 Feb;72(Pt 2):439–442. doi: 10.1099/0022-1317-72-2-439. [DOI] [PubMed] [Google Scholar]
  40. Williams M. A., Fukuda M. Accumulation of membrane glycoproteins in lysosomes requires a tyrosine residue at a particular position in the cytoplasmic tail. J Cell Biol. 1990 Sep;111(3):955–966. doi: 10.1083/jcb.111.3.955. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Wong S. H., Hong W. The SXYQRL sequence in the cytoplasmic domain of TGN38 plays a major role in trans-Golgi network localization. J Biol Chem. 1993 Oct 25;268(30):22853–22862. [PubMed] [Google Scholar]
  42. von Heijne G. Proline kinks in transmembrane alpha-helices. J Mol Biol. 1991 Apr 5;218(3):499–503. doi: 10.1016/0022-2836(91)90695-3. [DOI] [PubMed] [Google Scholar]

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