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Volume 270, Number 12, Issue of March 24, 1995 pp. 6523-6530
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
The Conserved Box 1 Motif of Cytokine Receptors Is Required for Association with JAK Kinases (*)

(Received for publication, November 21, 1994; and in revised form, January 18, 1995)

J. William Tanner (1), (§),  Wen Chen (1),  Robert L. Young (1),  Gregory D. Longmore (2),  (4),  Andrey S. Shaw (1)(¶)

From the  (1)Center for Immunology and the Departments of Pathology, (2)Medicine, and (3) (4)Cell Biology, Washington University School of Medicine, St. Louis, Missouri 63110

ABSTRACT

The JAK2 tyrosine kinase is known to associate with the receptors for growth hormone (GH) and erythropoietin (EPO) and with the interleukin-6 receptor signal transducing protein, gp130. Here we demonstrate that chimeric cytokine receptors which contain the cytoplasmic domain of the receptors for GH and EPO or for gp130 can form complexes with JAK2 when transiently co-expressed in HeLa cells. Mutational analyses of chimeras for the the GH and EPO receptors and gp130 demonstrated that box 1, a motif critical for cytokine receptor signal transduction, was required for the association of JAK2. Although JAK2 was capable of associating with all three of the chimeras, JAK1 co-precipitated only with the gp130 chimera. Association of JAK1 and JAK2 with cytokine receptor proteins, therefore, requires the highly conserved box 1 domain, but other sequences within the receptor proteins may influence the specificity of JAK binding. Mutational analysis of JAK2 revealed that multiple or complex protein sequences within JAK2 are required for association with cytokine receptors.

FOOTNOTES

*
This work was supported by a grant from Pfizer Central Research, Inc. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore by hereby marked ``advertisement'' in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

§
Recipient of a National Research Service Award from the National Institutes of Health.

To whom correspondence should be addressed: Center for Immunology, Dept. of Pathology, Box 8118, Washington University School of Medicine, 660 South Euclid, St. Louis, MO 63110. Tel.: 314-362-4614; Fax: 314-362-8888.

(^1)
The abbreviations used are: GH, growth hormone; GH-R, growth hormone receptor; EPO, erythropoietin; EPO-R, erythropoietin receptor; IL, interleukin; VSV G, vesicular stomatitis virus glycoprotein; PCR, polymerase chain reaction.

(^2)
W. Chen, J. W. Tanner, and A. S. Shaw, unpublished data.

ACKNOWLEDGEMENTS

We thank Dr. Robert Schreiber for helpful comments, discussions, and critical reading of the manuscript and Nick Staten for his encouragement and support during the early phases of these studies.

©1995 by The American Society for Biochemistry and Molecular Biology, Inc.

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