Raf Kinase Inhibitor Protein Interacts with NF-{kappa}B-Inducing Kinase and TAK1 and Inhibits NF-{kappa}B Activation

doi:10.1128/MCB.21.21.7207-7217.2001

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Molecular and Cellular Biology, November 2001, p. 7207-7217, Vol. 21, No. 21
0270-7306/01/$04.00+0 DOI: 10.1128/MCB.21.21.7207-7217.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Raf Kinase Inhibitor Protein Interacts with NF- $kappa$ B-Inducing Kinase and TAK1 and Inhibits NF- $kappa$ B Activation

Kam C. Yeung,¹^,^* David W. Rose,² Amardeep S. Dhillon,³ Diane Yaros,¹^, Marcus Gustafsson,¹ Devasis Chatterjee,¹ Brian McFerran,³ James Wyche,¹ Walter Kolch,³ and John M. Sedivy¹

Department of Molecular Biology, Cell Biology, and Biochemistry, Brown University, Providence, Rhode Island 02912¹; Department of Medicine and Whittier Diabetes Program, University of California $---$ San Diego, La Jolla, California 92093-0673²; and Beatson Institute for Cancer Research, CRC Beatson Laboratories, Bearsden, Glasgow G61 1BD, United Kingdom³

Received 8 January 2001/Returned for modification 14 March 2001/Accepted 2 August 2001

The Raf kinase inhibitor protein (RKIP) acts as a negative regulator of the mitogen-activated protein (MAP) kinase (MAPK)cascade initiated by Raf-1. RKIP inhibits the phosphorylationof MAP/extracellular signal-regulated kinase 1 (MEK1) by Raf-1by disrupting the interaction between these two kinases. We showhere that RKIP also antagonizes the signal transduction pathwaysthat mediate the activation of the transcription factor nuclearfactor kappa B (NF- $kappa$ B) in response to stimulation with tumor necrosisfactor alpha (TNF- $alpha$ ) or interleukin 1 beta. Modulation of RKIPexpression levels affected NF- $kappa$ B signaling independent of theMAPK pathway. Genetic epistasis analysis involving the ectopicexpression of kinases acting in the NF- $kappa$ B pathway indicated thatRKIP acts upstream of the kinase complex that mediates the phosphorylationand inactivation of the inhibitor of NF- $kappa$ B (I $kappa$ B). In vitro kinaseassays showed that RKIP antagonizes the activation of the I $kappa$ Bkinase (IKK) activity elicited by TNF- $alpha$ . RKIP physically interactedwith four kinases of the NF- $kappa$ B activation pathway, NF- $kappa$ B-inducingkinase, transforming growth factor beta-activated kinase 1, IKK $alpha$ ,and IKK $beta$ . This mode of action bears striking similarities to theinteractions of RKIP with Raf-1 and MEK1 in the MAPK pathway.Emerging data from diverse organisms suggest that RKIP and RKIP-relatedproteins represent a new and evolutionarily highly conserved familyof protein kinase regulators. Since the MAPK and NF- $kappa$ B pathwayshave physiologically distinct roles, the function of RKIP maybe, in part, to coordinate the regulation of thesepathways.

^* Corresponding author. Present address: Medical College of Ohio, Department of Biochemistry & Molecular Biology, 3035 ArlingtonAve., Toledo, OH 43614-5804. Phone: (419) 383-6658. Fax: (419)383-6228. E-mail: kyeung{at}mco.edu.

Present address: Rhode Island Hospital, Providence, RI02903.

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Raf Kinase Inhibitor Protein Interacts with NF-B-Inducing Kinase and TAK1 and Inhibits NF-B Activation

Raf Kinase Inhibitor Protein Interacts with NF- $kappa$ B-Inducing Kinase and TAK1 and Inhibits NF- $kappa$ B Activation