The human IL-11 receptor requires gp130 for signalling: demonstration by molecular cloning of the receptor.
We describe the molecular cloning of a cDNA for the alpha chain of the human IL-11 receptor (IL-11R alpha) and demonstrate the requirement of either the human or mouse gp130 molecule for signalling. cDNA clones encoding IL-11R alpha were isolated from a bone marrow cDNA library using a fragment from the murine IL-11R alpha as a probe. The human receptor was predicted to consist of 422 amino acids and was found to share 84% identity with the murine protein. In the extra-cellular region it exhibited a single hemopoietin domain with conserved cysteine residues and WSTWS motif. The transmembrane region was followed by a short cytoplasmic tail which did not contain a tyrosine kinase domain. Interaction of the human IL-11R alpha with murine gp130 was demonstrated: expression of the human IL-11R alpha in murine M1 cells which constitutively express murine gp130 (and murine LIF receptor), resulted in the generation of specific high-affinity binding sites for IL-11 (Kd = 250 pM). In addition, expression of the human IL-11R alpha in these cells permitted the induction of macrophage differentiation in response to IL-11. These results suggested that the human IL-11R alpha chain was able to form a functional receptor complex in association with murine gp130. The requirement of gp130 for signalling was confirmed by expression of the human IL-11R alpha in Ba/F3 cells. BaF3 cells that expressed the human IL-11R alpha alone showed binding of radiolabelled IL-11 but no proliferative response. Introduction of human gp130 into these cells resulted in high-affinity IL-11 binding sites and IL-11 dependent cellular proliferation. Thus these results demonstrated the absolute requirement of gp130 for signalling.
Nandurkar
H H
HH
Walter and Eliza Hall Institute of Medical Research, Victoria, Australia.
Hilton
D J
DJ
Nathan
P
P
Willson
T
T
Nicola
N
N
Begley
C G
CG
D000595:Amino Acid Sequence; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism*; D001483:Base Sequence; D015139:Blotting, Southern; D002455:Cell Division / Q000187:drug effects; D003001:Cloning, Molecular; D050822:Cytokine Receptor gp130; D006131:Growth Inhibitors; D016298:Hematopoietic Cell Growth Factors / Q000737:chemistry; D006801:Humans; D017370:Interleukin-11 / Q000494:pharmacology; D053703:Interleukin-11 Receptor alpha Subunit; D015850:Interleukin-6; D053605:Leukemia Inhibitory Factor; D053680:Leukemia Inhibitory Factor Receptor alpha Subunit; D008222:Lymphokines; D008562:Membrane Glycoproteins / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D018121:Receptors, Cytokine / Q000096:biosynthesis / Q000378:metabolism; D018123:Receptors, Interleukin / Q000096:biosynthesis / Q000737:chemistry / Q000502:physiology*; D053702:Receptors, Interleukin-11; D053606:Receptors, OSM-LIF; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D015398:Signal Transduction; D014162:Transfection
D015703:Antigens, CD; D006131:Growth Inhibitors; D016298:Hematopoietic Cell Growth Factors; C508428:IL11RA protein, human; C494199:IL6ST protein, human; C508429:Il11ra1 protein, mouse; C508431:Il11ra2 protein, mouse; C494200:Il6st protein, mouse; D017370:Interleukin-11; D053703:Interleukin-11 Receptor alpha Subunit; D015850:Interleukin-6; C507837:LIF protein, human; C507840:LIFR protein, human; D053605:Leukemia Inhibitory Factor; D053680:Leukemia Inhibitory Factor Receptor alpha Subunit; C507838:Lif protein, mouse; C507841:Lifr protein, mouse; D008222:Lymphokines; D008562:Membrane Glycoproteins; D018121:Receptors, Cytokine; D018123:Receptors, Interleukin; D053702:Receptors, Interleukin-11; D053606:Receptors, OSM-LIF; D011994:Recombinant Proteins; D050822:Cytokine Receptor gp130