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Molecular and Cellular Biology, October 2001, p. 6640-6650, Vol. 21, No. 19
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.19.6640-6650.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

B-Cell Receptor- and Phorbol Ester-Induced NF-kappa B and c-Jun N-Terminal Kinase Activation in B Cells Requires Novel Protein Kinase C's

Daniel Krappmann,* Alina Patke, Vigo Heissmeyer,dagger and Claus Scheidereit

Max-Delbrück-Centrum for Molecular Medicine, 13125 Berlin, Germany

Received 23 March 2001/Returned for modification 30 May 2001/Accepted 2 July 2001

Antigen receptor signaling is known to activate NF-kappa B in lymphocytes. While T-cell-receptor-induced NF-kappa B activation critically depends on novel protein kinase C theta  (PKCtheta ), the role of novel PKCs in B-cell stimulation has not been elucidated. In primary murine splenic B cells, we found high expression of the novel PKCs delta  and varepsilon  but only weak expression of the theta  isoform. Rottlerin blocks phorbol ester (phorbol myristate acetate [PMA])- or B-cell receptor (BCR)-mediated NF-kappa B and c-Jun N-terminal kinase (JNK) activation in primary B and T cells to a similar extent, suggesting that novel PKCs are positive regulators of signaling in hematopoietic cells. Mouse 70Z/3 pre-B cells have been widely used as a model for NF-kappa B activation in B cells. Similar to the situation in splenic B cells, rottlerin inhibits BCR and PMA stimulation of NF-kappa B in 70Z/3 cells. A derivative of 70Z/3 cells, 1.3E2 cells, are defective in NF-kappa B activation due to the lack of the Ikappa B kinase (IKKgamma ) protein. Ectopic expression of IKKgamma can rescue NF-kappa B activation in response to lipopolysaccharides (LPS) and interleukin-1beta (IL-1beta ), but not to PMA. In addition, PMA-induced activation of the mitogen-activated protein kinase JNK is blocked in 1.3E2 cells, suggesting that an upstream component common to both pathways is either missing or mutated. Analysis of various PKC isoforms revealed that exclusively PKCtheta was absent in 1.3E2 cells while it was expressed in 70Z/3 cells. Stable expression of either novel PKCtheta or -delta but not classical PKCbeta II in 1.3E2 IKKgamma -expressing cells rescues PMA activation of NF-kappa B and JNK signaling, demonstrating a critical role of novel PKCs for B-cell activation.

* Corresponding author. Mailing address: Max-Delbrück-Centrum for Molecular Medicine, Robert-Rössle Str. 10, 13125 Berlin, Germany. Phone: 49-30-9406-3751. Fax: 49-30-9406-3866. E-mail: dkrapp{at}mdc-berlin.de.

dagger Present address: Center for Blood Research, Harvard Medical School, Boston, MA 02115.

Molecular and Cellular Biology, October 2001, p. 6640-6650, Vol. 21, No. 19
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.19.6640-6650.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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