Dual role of the Jak1 FERM and kinase domains in cytokine receptor binding and in stimulation-dependent Jak activation.
Jak1 is a tyrosine kinase that noncovalently forms tight complexes with a variety of cytokine receptors and is critically involved in signal transduction via cytokines. Jaks are predicted to have a 4.1, ezrin, radixin, moesin (FERM) domain at their N terminus. FERM domains are composed of three structurally unrelated subdomains (F1, F2, and F3) which are in close contact to one another and form the clover-shaped FERM domain. We generated a model structure of the Jak1 FERM domain, based on solved FERM structures and the alignments with other FERM domains. To destabilize different subdomains and to uncover their exact function, we mutated specific hydrophobic residues conserved in FERM domains and involved in hydrophobic core interactions. In this study, we show that the structural integrity of the F2 subdomain of the FERM domain of Jak1 is necessary to bind the IFN-gammaRalpha. By mutagenesis of hydrophobic residues in the hydrophobic core between the three FERM subdomains, we find that the structural context of the FERM domain is necessary for the inhibition of Jak1 phosphorylation. Thus, FERM domain mutations can have repercussions on Jak1 function. Interestingly, a mutation in the kinase domain (Jak1-K907E), known to abolish the catalytic activity, also leads to an impaired binding to the IFN-gammaRalpha when this mutant is expressed at endogenous levels in U4C cells. Our data show that the structural integrity of both the FERM domain and of the kinase domain is essential for both receptor binding and catalytic function/autoinhibition.
Haan
Serge
S
Life Science Research Unit, Faculté des Sciences, de la Technologie et de la Communication, Université du Luxembourg, Luxembourg.
Margue
Christiane
C
Engrand
Arnaud
A
Rolvering
Catherine
C
Schmitz-Van de Leur
Hildegard
H
Heinrich
Peter C
PC
Behrmann
Iris
I
Haan
Claude
C
D000595:Amino Acid Sequence; D006801:Humans; D007371:Interferon-gamma / Q000378:metabolism; D053613:Janus Kinase 1 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D009154:Mutation; D010766:Phosphorylation; D017434:Protein Structure, Tertiary / Q000235:genetics; D018121:Receptors, Cytokine / Q000378:metabolism*; D017471:Receptors, Interferon / Q000378:metabolism*; D000096743:Interferon gamma Receptor
D007371:Interferon-gamma; D053613:Janus Kinase 1; D018121:Receptors, Cytokine; D017471:Receptors, Interferon; D000096743:Interferon gamma Receptor