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Updated: May 10, 2026

High-throughput Quantitative Real-time RT-PCR Assay for Determining Expression Profiles of Types I and III Interferon Subtypes
10:00

High-throughput Quantitative Real-time RT-PCR Assay for Determining Expression Profiles of Types I and III Interferon Subtypes

Published on: March 24, 2015

Ligand-independent interaction of the type I interferon receptor complex is necessary to observe its biological

Christopher D Krause1, Gina Digioia, Lara S Izotova

  • 1Department of Biochemistry and Molecular Biology, Robert Wood Johnson Medical School, The University of Medicine and Dentistry of New Jersey, 675 Hoes Lane West, Piscataway, NJ 08855, USA.

Cytokine
|July 9, 2013
PubMed

Ectopic coexpression of the two chains of the Type I and Type III interferon (IFN) receptor complexes (IFN-αR1 and IFN-αR2c, or IFN-λR1 and IL-10R2) yielded sensitivity to IFN-alpha or IFN-lambda in only some cells. We found that IFN-αR1 and IFN-αR2c exhibit FRET only when expressed at equivalent and low levels. Expanded clonal cell lines expressing both IFN-αR1 and IFN-αR2c were sensitive to IFN-alpha only when IFN-αR1 and IFN-αR2c exhibited FRET in the absence of human IFN-alpha. Coexpression of RACK-1 or Jak1 enhanced the affinity of the interaction between IFN-αR1 and IFN-αR2c. Both IFN-αR1 and IFN-αR2c exhibited FRET with Jak1 and Tyk2. Together with data showing that disruption of the preassociation between the IFN-gamma receptor chains inhibited its biological activity, we propose that biologically active IFN receptors require ligand-independent juxtaposition of IFN receptor chains assisted by their associated cytosolic proteins.

Keywords:
CiFPConfocal fluorescence spectroscopyECFPEGFPEYFPFRETIFNIFN-αIFN-γIFN-λInterferon receptorOFPRACK-1Receptor reconstitutionStFPenhanced citrine fluorescent proteinenhanced cyan fluorescent proteinenhanced green fluorescent proteinenhanced yellow fluorescent proteinfluorescence resonance energy transferinterferoninterferon-alphainterferon-gammainterferon-lambdaorange fluorescent proteinreceptor for activated protein kinase-1strawberry fluorescent protein

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