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Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1995 Aug;15(8):4208–4214. doi: 10.1128/mcb.15.8.4208

Ligand-induced association of the type I interferon receptor components.

B Cohen 1, D Novick 1, S Barak 1, M Rubinstein 1
PMCID: PMC230659  PMID: 7623815

Abstract

Two transmembrane polypeptides, IFNAR and IFN-alpha/Beta R, were previously identified as essential components of the type I interferon (IFN) receptor, but their interrelationship and role in ligand binding were not clear. To study these issues, we stably expressed and characterized the two polypeptides in host murine cells. In human cells, native IFN-alpha/beta R is a 102-kDa protein but upon reduction only a 51-kDa protein is detected. In host murine cells human IFN-alpha/beta R was expressed as a 51-kDa protein. Host cells expressing IFN-alpha/beta R bound IFN-alpha 2 with a high affinity (Kd of 3.6 nM), whereas cells expressing IFNAR exhibited no ligand binding. Upon coexpression of IFNAR and the 51-kDa IFN-alpha/beta R, the affinity for IFN-alpha 2 was increased 10-fold, approaching that of the native receptor. We show by cross-linking that both the cloned (51-kDa) and native (102-kDa) IFN-alpha/beta R bind IFN-alpha 2 to form an intermediate product, while IFNAR associates with this product to form a ternary complex. Hence, IFNAR and IFN-alpha/beta R are components of a common type I IFN receptor, cooperating in ligand binding. Ligand-induced association of IFNAR and IFN-alpha/beta R probably triggers transmembrane signaling.

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Selected References

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  1. Abramovich C., Shulman L. M., Ratovitski E., Harroch S., Tovey M., Eid P., Revel M. Differential tyrosine phosphorylation of the IFNAR chain of the type I interferon receptor and of an associated surface protein in response to IFN-alpha and IFN-beta. EMBO J. 1994 Dec 15;13(24):5871–5877. doi: 10.1002/j.1460-2075.1994.tb06932.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barbieri G., Velazquez L., Scrobogna M., Fellous M., Pellegrini S. Activation of the protein tyrosine kinase tyk2 by interferon alpha/beta. Eur J Biochem. 1994 Jul 15;223(2):427–435. doi: 10.1111/j.1432-1033.1994.tb19010.x. [DOI] [PubMed] [Google Scholar]
  3. Boshart M., Klüppel M., Schmidt A., Schütz G., Luckow B. Reporter constructs with low background activity utilizing the cat gene. Gene. 1992 Jan 2;110(1):129–130. doi: 10.1016/0378-1119(92)90456-y. [DOI] [PubMed] [Google Scholar]
  4. Branca A. A., Baglioni C. Evidence that types I and II interferons have different receptors. Nature. 1981 Dec 24;294(5843):768–770. doi: 10.1038/294768a0. [DOI] [PubMed] [Google Scholar]
  5. Cohen B., Peretz D., Vaiman D., Benech P., Chebath J. Enhancer-like interferon responsive sequences of the human and murine (2'-5') oligoadenylate synthetase gene promoters. EMBO J. 1988 May;7(5):1411–1419. doi: 10.1002/j.1460-2075.1988.tb02958.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Colamonici O. R., Domanski P. Identification of a novel subunit of the type I interferon receptor localized to human chromosome 21. J Biol Chem. 1993 May 25;268(15):10895–10899. [PubMed] [Google Scholar]
  7. Colamonici O., Yan H., Domanski P., Handa R., Smalley D., Mullersman J., Witte M., Krishnan K., Krolewski J. Direct binding to and tyrosine phosphorylation of the alpha subunit of the type I interferon receptor by p135tyk2 tyrosine kinase. Mol Cell Biol. 1994 Dec;14(12):8133–8142. doi: 10.1128/mcb.14.12.8133. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Constantinescu S. N., Croze E., Wang C., Murti A., Basu L., Mullersman J. E., Pfeffer L. M. Role of interferon alpha/beta receptor chain 1 in the structure and transmembrane signaling of the interferon alpha/beta receptor complex. Proc Natl Acad Sci U S A. 1994 Sep 27;91(20):9602–9606. doi: 10.1073/pnas.91.20.9602. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Eid P., Mogensen K. E. Isolated interferon alpha-receptor complexes stabilized in vitro. FEBS Lett. 1983 May 30;156(1):157–160. doi: 10.1016/0014-5793(83)80268-3. [DOI] [PubMed] [Google Scholar]
  10. Frohman M. A., Dush M. K., Martin G. R. Rapid production of full-length cDNAs from rare transcripts: amplification using a single gene-specific oligonucleotide primer. Proc Natl Acad Sci U S A. 1988 Dec;85(23):8998–9002. doi: 10.1073/pnas.85.23.8998. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Joshi A. R., Sarkar F. H., Gupta S. L. Interferon receptors. Cross-linking of human leukocyte interferon alpha-2 to its receptor on human cells. J Biol Chem. 1982 Dec 10;257(23):13884–13887. [PubMed] [Google Scholar]
  13. Kishimoto T., Taga T., Akira S. Cytokine signal transduction. Cell. 1994 Jan 28;76(2):253–262. doi: 10.1016/0092-8674(94)90333-6. [DOI] [PubMed] [Google Scholar]
  14. Lim J. K., Langer J. A. Cloning and characterization of a bovine alpha interferon receptor. Biochim Biophys Acta. 1993 Jun 25;1173(3):314–319. doi: 10.1016/0167-4781(93)90129-2. [DOI] [PubMed] [Google Scholar]
  15. Mizushima S., Nagata S. pEF-BOS, a powerful mammalian expression vector. Nucleic Acids Res. 1990 Sep 11;18(17):5322–5322. doi: 10.1093/nar/18.17.5322. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Munson P. J., Rodbard D. Ligand: a versatile computerized approach for characterization of ligand-binding systems. Anal Biochem. 1980 Sep 1;107(1):220–239. doi: 10.1016/0003-2697(80)90515-1. [DOI] [PubMed] [Google Scholar]
  17. Müller M., Briscoe J., Laxton C., Guschin D., Ziemiecki A., Silvennoinen O., Harpur A. G., Barbieri G., Witthuhn B. A., Schindler C. The protein tyrosine kinase JAK1 complements defects in interferon-alpha/beta and -gamma signal transduction. Nature. 1993 Nov 11;366(6451):129–135. doi: 10.1038/366129a0. [DOI] [PubMed] [Google Scholar]
  18. Müller U., Steinhoff U., Reis L. F., Hemmi S., Pavlovic J., Zinkernagel R. M., Aguet M. Functional role of type I and type II interferons in antiviral defense. Science. 1994 Jun 24;264(5167):1918–1921. doi: 10.1126/science.8009221. [DOI] [PubMed] [Google Scholar]
  19. Nguyen V. T., Morange M., Bensaude O. Firefly luciferase luminescence assays using scintillation counters for quantitation in transfected mammalian cells. Anal Biochem. 1988 Jun;171(2):404–408. doi: 10.1016/0003-2697(88)90505-2. [DOI] [PubMed] [Google Scholar]
  20. Nordeen S. K. Luciferase reporter gene vectors for analysis of promoters and enhancers. Biotechniques. 1988 May;6(5):454–458. [PubMed] [Google Scholar]
  21. Novick D., Cohen B., Rubinstein M. Soluble interferon-alpha receptor molecules are present in body fluids. FEBS Lett. 1992 Dec 21;314(3):445–448. doi: 10.1016/0014-5793(92)81523-o. [DOI] [PubMed] [Google Scholar]
  22. Novick D., Cohen B., Rubinstein M. The human interferon alpha/beta receptor: characterization and molecular cloning. Cell. 1994 May 6;77(3):391–400. doi: 10.1016/0092-8674(94)90154-6. [DOI] [PubMed] [Google Scholar]
  23. Pestka S., Langer J. A., Zoon K. C., Samuel C. E. Interferons and their actions. Annu Rev Biochem. 1987;56:727–777. doi: 10.1146/annurev.bi.56.070187.003455. [DOI] [PubMed] [Google Scholar]
  24. Rubinstein S., Familletti P. C., Pestka S. Convenient assay for interferons. J Virol. 1981 Feb;37(2):755–758. doi: 10.1128/jvi.37.2.755-758.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Silvennoinen O., Ihle J. N., Schlessinger J., Levy D. E. Interferon-induced nuclear signalling by Jak protein tyrosine kinases. Nature. 1993 Dec 9;366(6455):583–585. doi: 10.1038/366583a0. [DOI] [PubMed] [Google Scholar]
  26. Sompayrac L. M., Danna K. J. Efficient infection of monkey cells with DNA of simian virus 40. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7575–7578. doi: 10.1073/pnas.78.12.7575. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Ullrich A., Schlessinger J. Signal transduction by receptors with tyrosine kinase activity. Cell. 1990 Apr 20;61(2):203–212. doi: 10.1016/0092-8674(90)90801-k. [DOI] [PubMed] [Google Scholar]
  28. Uzé G., Di Marco S., Mouchel-Vielh E., Monneron D., Bandu M. T., Horisberger M. A., Dorques A., Lutfalla G., Mogensen K. E. Domains of interaction between alpha interferon and its receptor components. J Mol Biol. 1994 Oct 21;243(2):245–257. doi: 10.1006/jmbi.1994.1651. [DOI] [PubMed] [Google Scholar]
  29. Uzé G., Lutfalla G., Gresser I. Genetic transfer of a functional human interferon alpha receptor into mouse cells: cloning and expression of its cDNA. Cell. 1990 Jan 26;60(2):225–234. doi: 10.1016/0092-8674(90)90738-z. [DOI] [PubMed] [Google Scholar]

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