Germinal Center Kinase Is Required for Optimal Jun N-Terminal Kinase Activation by Toll-Like Receptor Agonists and Is Regulated by the Ubiquitin Proteasome System and Agonist-Induced, TRAF6-Dependent Stabilization

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Published inMolecular and Cellular Biology Vol. 24; no. 20; pp. 9165 - 9175
Main Authors Zhong, Jian, Kyriakis, John M.
Format Journal Article
LanguageEnglish
Published United States American Society for Microbiology 01.10.2004
Taylor & Francis
Subjects
Online AccessGet full text
ISSN0270-7306
1098-5549
1067-8824
1098-5549
DOI10.1128/MCB.24.20.9165-9175.2004

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Germinal center kinase (GCK), a member of the Ste20 family, selectively activates the Jun N-terminal kinase (JNK) group of mitogen-activated protein kinases. Here, we show that endogenous GCK is activated by polyinosine-polycytidine [poly(IC)] and lipopolysaccharides (LPS), lipid A, interleukin-1 (IL-1), and engagement of CD40, all agonists that require TRAF6 for JNK activation. RNA interference experiments indicate that GCK is required for the maximal activation of JNK by LPS, lipid A, poly(IC), and, to a lesser extent, IL-1 and engagement of CD40. GCK is ubiquitinated in situ and stabilized by inhibitors of the proteasome, indicating that GCK is subject to proteasomal turnover. GCK is constitutively active, and the kinase activity of GCK is required for GCK ubiquitination. Agonist activation of GCK involves the TRAF6-dependent transient stabilization of the GCK polypeptide rather than an increase in intrinsic kinase activity. Our results identify a physiologic function and unexpected mode of regulation for GCK.
Germinal center kinase (GCK), a member of the Ste20 family, selectively activates the Jun N-terminal kinase (JNK) group of mitogen-activated protein kinases. Here, we show that endogenous GCK is activated by polyinosine-polycytidine [poly(IC)] and lipopolysaccharides (LPS), lipid A, interleukin-1 (IL-1), and engagement of CD40, all agonists that require TRAF6 for JNK activation. RNA interference experiments indicate that GCK is required for the maximal activation of JNK by LPS, lipid A, poly(IC), and, to a lesser extent, IL-1 and engagement of CD40. GCK is ubiquitinated in situ and stabilized by inhibitors of the proteasome, indicating that GCK is subject to proteasomal turnover. GCK is constitutively active, and the kinase activity of GCK is required for GCK ubiquitination. Agonist activation of GCK involves the TRAF6-dependent transient stabilization of the GCK polypeptide rather than an increase in intrinsic kinase activity. Our results identify a physiologic function and unexpected mode of regulation for GCK.Germinal center kinase (GCK), a member of the Ste20 family, selectively activates the Jun N-terminal kinase (JNK) group of mitogen-activated protein kinases. Here, we show that endogenous GCK is activated by polyinosine-polycytidine [poly(IC)] and lipopolysaccharides (LPS), lipid A, interleukin-1 (IL-1), and engagement of CD40, all agonists that require TRAF6 for JNK activation. RNA interference experiments indicate that GCK is required for the maximal activation of JNK by LPS, lipid A, poly(IC), and, to a lesser extent, IL-1 and engagement of CD40. GCK is ubiquitinated in situ and stabilized by inhibitors of the proteasome, indicating that GCK is subject to proteasomal turnover. GCK is constitutively active, and the kinase activity of GCK is required for GCK ubiquitination. Agonist activation of GCK involves the TRAF6-dependent transient stabilization of the GCK polypeptide rather than an increase in intrinsic kinase activity. Our results identify a physiologic function and unexpected mode of regulation for GCK.
Author Jian Zhong
John M. Kyriakis
AuthorAffiliation Molecular Cardiology Research Institute and Department of Medicine, Tufts-New England Medical Center, and Department of Medicine, Tufts University School of Medicine, Boston, Massachusetts
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Corresponding author. Mailing address: Molecular Cardiology Research Institute Tufts-New England Medical Center, 750 Washington St., Box 8486, Boston, MA 02111. Phone: (617) 636-5190. Fax: (617) 636-5204. E-mail: jkyriakis@tufts-nemc.org.
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Germinal center kinase (GCK), a member of the Ste20 family, selectively activates the Jun N-terminal kinase (JNK) group of mitogen-activated protein kinases....
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StartPage 9165
SubjectTerms Animals
CD40 Antigens - metabolism
Cell Growth and Development
Cell Line
Enzyme Activation
Humans
Interferon Inducers - metabolism
Interleukin-1 - metabolism
JNK Mitogen-Activated Protein Kinases - metabolism
Lipid A - metabolism
Lipopolysaccharides - metabolism
MAP Kinase Kinase Kinase 1 - metabolism
Membrane Glycoproteins - agonists
Membrane Glycoproteins - metabolism
Poly I-C - metabolism
Proteasome Endopeptidase Complex - metabolism
Proteasome Inhibitors
Protein-Serine-Threonine Kinases - genetics
Protein-Serine-Threonine Kinases - metabolism
Receptors, Cell Surface - agonists
Receptors, Cell Surface - metabolism
RNA Interference
Signal Transduction - physiology
TNF Receptor-Associated Factor 6 - metabolism
Toll-Like Receptors
Ubiquitin - metabolism
Title Germinal Center Kinase Is Required for Optimal Jun N-Terminal Kinase Activation by Toll-Like Receptor Agonists and Is Regulated by the Ubiquitin Proteasome System and Agonist-Induced, TRAF6-Dependent Stabilization
URI http://mcb.asm.org/content/24/20/9165.abstract
https://www.tandfonline.com/doi/abs/10.1128/MCB.24.20.9165-9175.2004
https://www.ncbi.nlm.nih.gov/pubmed/15456887
https://www.proquest.com/docview/19721683
https://www.proquest.com/docview/66925827
https://pubmed.ncbi.nlm.nih.gov/PMC517887
http://doi.org/10.1128/MCB.24.20.9165-9175.2004
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