Distinct domains of the protein tyrosine kinase tyk2 required for binding of interferon-alpha/beta and for signal transduction.
tyk2 belongs to the JAK family of nonreceptor protein tyrosine kinases recently found implicated in signaling through a large number of cytokine receptors. These proteins are characterized by a large amino-terminal region and two tandemly arranged kinase domains, a kinase-like and a tyrosine kinase domain. Genetic and biochemical evidence supports the requirement for tyk2 in interferon-alpha/beta binding and signaling. To study the role of the distinct domains of tyk2, constructs lacking one or both kinase domains were stably transfected in recipient cells lacking the endogenous protein. Removal of either or both kinase domains resulted in loss of the in vitro kinase activity. The mutant form truncated of the tyrosine kinase domain was found to reconstitute binding of interferon-alpha 8 and partial signaling. While no contribution of this protein toward interferon-beta binding was evident, increased signaling could be measured. The mutant form lacking both kinase domains did not exhibit any detectable activity. Altogether, these results show that a sequential deletion of domains engenders a sequential loss of function and that the different domains of tyk2 have distinct functions, all essential for full interferon-alpha and -beta binding and signaling.
Velazquez
L
L
Institut Pasteur, INSERM U 276, Paris, France.
Mogensen
K E
KE
Barbieri
G
G
Fellous
M
M
Uzé
G
G
Pellegrini
S
S
D000818:Animals; D001665:Binding Sites; D015152:Blotting, Northern; D002460:Cell Line; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D015151:Immunoblotting; D007370:Interferon Type I / Q000494:pharmacology; D016898:Interferon-alpha / Q000378:metabolism*; D016899:Interferon-beta / Q000378:metabolism* / Q000494:pharmacology; D007700:Kinetics; D016296:Mutagenesis; D010766:Phosphorylation; D019000:Phosphotyrosine; D014176:Protein Biosynthesis; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D012333:RNA, Messenger / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D015183:Restriction Mapping; D017384:Sequence Deletion; D015398:Signal Transduction; D053634:TYK2 Kinase; D014162:Transfection; D014443:Tyrosine / Q000031:analogs & derivatives / Q000032:analysis / Q000378:metabolism
D007370:Interferon Type I; D016898:Interferon-alpha; D011506:Proteins; D012333:RNA, Messenger; D011994:Recombinant Proteins; D019000:Phosphotyrosine; D014443:Tyrosine; D016899:Interferon-beta; D011505:Protein-Tyrosine Kinases; D053634:TYK2 Kinase; C070139:TYK2 protein, human