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J. Biol. Chem., Vol. 276, Issue 25, 22476-22484, June 22, 2001
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From Ciliary neurotrophic factor (CNTF) is a cytokine
supporting the differentiation and survival of a number of neural cell
types. Its receptor complex consists of a ligand-binding component,
CNTF receptor (CNTFR), associated with two signaling receptor
components, gp130 and leukemia inhibitory factor receptor (LIFR).
Striking phenotypic differences between CNTF- and CNTFR-deficient mice suggest that CNTFR serves as a receptor for a second developmentally important ligand. We recently demonstrated that cardiotrophin-like cytokine (CLC) associates with the soluble orphan receptor
cytokine-like factor-1 (CLF) to form a heterodimeric cytokine that
displayed activities only on cells expressing the tripartite CNTF
receptor on their surface. In this present study we examined the
membrane binding of the CLC/CLF composite cytokine and observed a
preferential interaction of the cytokine with the CNTFR subunit.
Signaling pathways recruited by the CLC/CLF complex in human
neuroblastoma cell lines were also analyzed in detail. The results
obtained showed an activation of Janus kinases (JAK1, JAK2, and TYK2)
leading to a tyrosine phosphorylation of the gp130 and LIFR. The
phosphorylated signaling receptors served in turn as docking proteins
for signal transducing molecules such as STAT3 and SHP-2. In
vitro analysis revealed that the gp130-LIFR pathway could also
stimulate the phosphatidylinositol 3-kinase and the
mitogen-activated protein kinase pathways. In contrast to that reported
before for CNTF, soluble CNTFR failed to promote the action CLC/CLF,
and an absolute requirement of the membrane form of CNTFR was required
to generate a functional response to the composite cytokine. This study
reinforces the functional similarity between CNTF and the CLC/CLF
composite cytokine defining the second ligand for CNTFR.
Signaling Pathways Recruited by the Cardiotrophin-like
Cytokine/Cytokine-like Factor-1 Composite Cytokine
SPECIFIC REQUIREMENT OF THE MEMBRANE-BOUND FORM OF CILIARY
NEUROTROPHIC FACTOR RECEPTOR 
COMPONENT*
§,¶,,,
,
INSERM EMI-9928, CHU d'Angers, 4 rue Larrey,
49003 Angers, France and the ** Centre d'Immunologie Pierre Fabre, 5 avenue Napoléon III,
74164 Saint Julien en Genevois, France
*
This work was supported by a grant from the Association
Française contre les Myopathies and the Ligue Nationale contre le Cancer.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Supported by a grant from the Département du Maine et Loire.
To whom correspondence should be addressed: INSERM E 9928, CHU d'Angers, 4 rue Larrey, 49033 Angers Cedex, France.
Tel.: 33-2-41-35-47-31; Fax: 33-2-41-73-16-30; E-mail:
hugues.gascan@univ-angers.fr.
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