Knockout and reconstitution of a functional human type I interferon receptor complex

• Published in: The Journal of biological chemistry Vol. 269; no. 29; pp. 18747 - 18749
• Main Authors: CLEARY, C. M, DONNELLY, R. J, JAEMOG SOH, MARIANO, T. M, PESTKA, S
• Format: Journal Article
• Language: English
• Published: Bethesda, MD American Society for Biochemistry and Molecular Biology 22.07.1994
• Subjects: Biological and medical sciences; Cell receptors; Cell structures and functions; Chromosomes, Artificial, Yeast; Fundamental and applied biological sciences. Psychology; Histocompatibility Antigens Class I - metabolism; Hormone receptors. Growth factor receptors. Cytokine receptors. Prostaglandin receptors; Humans; Interferon Type I - metabolism; Molecular and cellular biology; Mutagenesis, Insertional; Receptor, Interferon alpha-beta; Receptors, Interferon - genetics; Restriction Mapping; Sequence Deletion; Viral Interference; Human; Ligand binding; Alpha interferon; Major histocompatibility system; Cytokine; Reconstituted system; Antiviral; Mutation; Subunit; Structure function relationship; Biological receptor
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/s0021-9258(17)32231-7

The functional subunits of the human Type I interferon (IFN) receptor complex have not been defined. Using site-specific recombination in a yeast artificial chromosome (YAC), we have produced a deletion within the human IFN-alpha receptor (Hu-IFN-alpha R1) gene which eliminates exon II of the gene. This deletion effectively eliminates the MHC Class I antigen induction and antiviral activity previously reported for this fully functional parental YAC clone (Soh, J., Mariano, T. M., Lim, J.-K., Izotova, L., Mirochnitchenko, O., Schwartz, B., Langer, J., and Pestka, S. (1994c) J. Biol. Chem. 269, 18102-18110). We have successfully reconstituted this activity by expression of the cDNA encoding the Hu-IFN-alpha R1 component (Uzé, G., Lutfalla, G., and Gresser, I. (1990) Cell 60, 225-234) in cells containing the YAC with this deletion. The Hu-IFN-alpha R1 subunit thus plays a critical role in the functional human Type I IFN receptor complex, whose components are encoded on this YAC. In addition, as binding of ligands is retained in the cells containing the YAC with the deletion, it is clear a second subunit encoded on the YAC is responsible for ligand binding activity. This system will now allow the identification of additional subunits involved in the response to the Type I IFNs and the functional significance of each.