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J Biol Chem, Vol. 274, Issue 49, 34838-34845, December 3, 1999

Dimerization of the Interferon Type I Receptor IFNaR2-2 Is Sufficient for Induction of Interferon Effector Genes but Not for Full Antiviral Activity

Els Pattyn, Xaveer Van Ostade, Liesbeth Schauvliege, Annick Verhee, Michael Kalai^¶, Joël Vandekerckhove, and Jan Tavernier

From the  Flanders Interuniversity Institute for Biotechnology, Department of Medical Protein Research, Faculty of Medicine, and the ^¶ Department of Molecular Biology, Faculty of Sciences, University of Ghent, K. L. Ledeganckstraat 35, B-9000 Ghent, Belgium

We constructed chimeric receptors wherein the extracellular domain of the erythropoietin receptor (EpoR) was fused to the transmembrane and intracellular domains of the interferon (IFN) type I receptor subunits, IFNaR1 or IFNaR2-2. Transfection into 2fTGH and Tyk2-deficient 11,1 cells showed that EpoR/IFNaR2-2 alone was able to transduce a signal upon stimulation with erythropoietin (Epo), as judged by induction of the interferon type I-inducible 6-16 promoter. In contrast, protection against infection with encephalomyocarditis virus or vesicular stomatitis virus was reduced or absent, respectively. To further investigate the role of IFNaR1 in the induction of an antiviral state, we analyzed the Epo- versus IFN-induced transcription of a set of genes, involved in antiviral protection. Up to 24 h after stimulation with Epo or IFN, comparable transcription of the p56, dsRNA-dependent protein kinase, 2'-5'A synthetase, and MxA genes was seen. However, at later time points, only in the case of Epo induction, a sharp decrease of mRNA levels was observed. Western blotting analysis of dsRNA-dependent protein kinase showed a similar pattern at the protein level. Taken together, our results imply a role for IFNaR1 in the induction of sustained mRNA and protein levels that are likely required for optimal antiviral activity.

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