Knockout and reconstitution of a functional human type I interferon receptor complex.
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.
Department of Molecular Genetics and Microbiology, University of Medicine and Dentistry of New Jersey, Robert Wood Johnson Medical School, Piscataway 08854-5635.
C M
CM
Cleary
R J
RJ
Donnelly
J
J
Soh
T M
TM
Mariano
S
S
Pestka
D018244:Chromosomes, Artificial, Yeast; D015395:Histocompatibility Antigens Class I / Q000378:metabolism; D006801:Humans; D007370:Interferon Type I / Q000378:metabolism*; D016254:Mutagenesis, Insertional; D053633:Receptor, Interferon alpha-beta; D017471:Receptors, Interferon / Q000235:genetics*; D015183:Restriction Mapping; D017384:Sequence Deletion; D014762:Viral Interference
D015395:Histocompatibility Antigens Class I; D007370:Interferon Type I; D017471:Receptors, Interferon; D053633:Receptor, Interferon alpha-beta