title: The extracellular and transmembrane domains of the γC and interleukin (IL)-13 receptor α1 chains, not their cytoplasmic domains, dictate the nature of signaling responses to IL-4 and IL-13.
abstract: Previously, we demonstrated that the γC subunit of type I IL-4 receptor was required for robust tyrosine phosphorylation of the downstream adapter protein, IRS-2, correlating with the expression of genes (ArgI, Retnla, and Chi3l3) characteristic of alternatively activated macrophages. We located an I4R-like motif (IRS-2 docking sequence) in the γC cytoplasmic domain but not in the IL-13Rα1. Thus, we predicted that the γC tail directed enhanced IRS-2 phosphorylation. To test this, IL-4 signaling responses were examined in a mutant of the key I4R motif tyrosine residue (Y325F) and different γC truncation mutants (γ285, γ308, γ318, γ323, and γFULL LENGTH (FL)) co-expressed in L-cells or CHO cells with wild-type (WT) IL-4Rα. Surprisingly, IRS-1 phosphorylation was not diminished in Y325F L-cell mutants suggesting Tyr-325 was not required for the robust insulin receptor substrate response. IRS-2, STAT6, and JAK3 phosphorylation was observed in CHO cells expressing γ323 and γFL but not in γ318 and γ285 mutants. In addition, when CHO cells expressed γ318, γ323, or γFL with IL-2Rβ, IL-2 induced phospho-STAT5 only in the γ323 and γFL clones. Our data suggest that a smaller (5 amino acid) interval than previously determined is necessary for JAK3 activation/γC-mediated signaling in response to IL-4 and IL-2. Chimeric receptor chains of the γC tail fused to the IL-13Rα1 extracellular and transmembrane domain did not elicit robust IRS-2 phosphorylation in response to IL-13 suggesting that the extracellular/transmembrane domains of the IL-4/IL-13 receptor, not the cytoplasmic domains, control signaling efficiency. Understanding this pathway fully will lead to rational drug design for allergic disease.
authors: Center for Vascular and Inflammatory Diseases, University of Maryland Baltimore, Baltimore, Maryland 21201, USA.
authors: Nicola M
authors: NM
authors: Heller
authors: Xiulan
authors: X
authors: Qi
authors: Franck
authors: F
authors: Gesbert
authors: Achsah D
authors: AD
authors: Keegan
mesh_terms: D000818:Animals; D016466:CHO Cells; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D016207:Cytokines / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D005434:Flow Cytometry; D053631:Interleukin Receptor Common gamma Subunit / Q000737:chemistry*; D018793:Interleukin-13 / Q000737:chemistry; D053661:Interleukin-13 Receptor alpha1 Subunit / Q000737:chemistry*; D015847:Interleukin-4 / Q000737:chemistry*; D051379:Mice; D008954:Models, Biological; D009154:Mutation; D010766:Phosphorylation; D017434:Protein Structure, Tertiary; D017455:Receptors, IgE / Q000378:metabolism; D019948:Receptors, Interleukin-4 / Q000378:metabolism; D050800:STAT6 Transcription Factor / Q000378:metabolism; D015398:Signal Transduction
chemical_list: D016207:Cytokines; D053631:Interleukin Receptor Common gamma Subunit; D018793:Interleukin-13; D053661:Interleukin-13 Receptor alpha1 Subunit; D017455:Receptors, IgE; D019948:Receptors, Interleukin-4; D050800:STAT6 Transcription Factor; D015847:Interleukin-4