Division of Rheumatology, Allergy, and Immunology, UCSD School of Medicine, 9500 Gilman Drive, La Jolla, CA 92093Search for more papers by this author

Monisha Sundarrajan,

Monisha Sundarrajan

University of California, San Diego, School of Medicine, La Jolla

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David L. Boyle,

David L. Boyle

University of California, San Diego, School of Medicine, La Jolla

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Martine Chabaud-Riou,

Martine Chabaud-Riou

University of California, San Diego, School of Medicine, La Jolla

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Deepa Hammaker,

Deepa Hammaker

University of California, San Diego, School of Medicine, La Jolla

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Gary S. Firestein,

Corresponding Author

Gary S. Firestein

[email protected]

University of California, San Diego, School of Medicine, La Jolla

Division of Rheumatology, Allergy, and Immunology, UCSD School of Medicine, 9500 Gilman Drive, La Jolla, CA 92093Search for more papers by this author

First published: 11 September 2003

https://doi.org/10.1002/art.11228

Citations: 48

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Abstract

Objective

The mitogen-activated protein (MAP) kinase JNK is a key regulator of interleukin-1 (IL-1)–induced collagenase gene expression and joint destruction in arthritis. Two upstream kinases, MKK-4 and MKK-7, have been identified as potential activators of JNK. However, the role of MAP kinase kinases (MAPKKs) and their functional organization within fibroblast-like synoviocytes (FLS) have not been defined. We therefore evaluated the interactions between the various MAP kinase components and determined their subcellular localization.

Methods

MKKs were identified by immunohistochemistry of rheumatoid arthritis (RA) and osteoarthritis (OA) synovium. Western blotting was used to determine the expression of FLS. Immunoprecipitation experiments using antibodies specific for MKK-4, MKK-7, and JNK were performed. Phosphospecific antibodies and immunohistochemistry were used to evaluate the activation state of synovial MKK-4 and MKK-7. Confocal microscopy was used to determine the subcellular location of the kinases.

Results

Immunohistochemistry studies demonstrated abundant MKK-4 and MKK-7 in RA and OA synovium, but the levels of phosphorylated kinases were significantly higher in RA synovium. MKK-4 and MKK-7 were constitutively expressed by cultured RA and OA FLS, and IL-1 stimulation resulted in rapid phosphorylation of both kinases. JNK was detected in MKK-4 and MKK-7 immunoprecipitates. Furthermore, MKK-4 coprecipitated with MKK-7 and vice versa, indicating that the 3 kinases form a stable complex in FLS. Confocal microscopy confirmed that JNK, MKK-4, and MKK-7 colocalized in the cytoplasm, with JNK migrating to the nucleus after IL-1 stimulation. The signal complex containing MKK-4, MKK-7, and JNK was functionally active and able to phosphorylate c-Jun after IL-1 stimulation of FLS.

Conclusion

These studies demonstrate that JNK, MKK-4, and MKK-7 form an active signaling complex in FLS. This novel JNK signalsome is activated in response to IL-1 and migrates to the nucleus. The JNK signalsome represents a new target for therapeutic interventions designed to prevent joint destruction.

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Citing Literature

Volume48, Issue9

September 2003

Pages 2450-2460

Expression of the MAPK kinases MKK-4 and MKK-7 in rheumatoid arthritis and their role as key regulators of JNK

Abstract

Objective

Methods

Results

Conclusion

REFERENCES

Citing Literature

References

Information

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Expression of the MAPK kinases MKK-4 and MKK-7 in rheumatoid arthritis and their role as key regulators of JNK

Abstract

Objective

Methods

Results

Conclusion

REFERENCES

Citing Literature

References

Related

Information