Distinct subcellular localization and substrate specificity of extracellular signal-regulated kinase in B cells upon stimulation with IgM and CD40.
We and others previously observed that IgM and CD40 stimulation in murine B cells resulted in activation of extracellular signal-regulated kinase (ERK), a subfamily of mitogen-activated protein kinase. The present study demonstrated that ERK was rapidly phosphorylated and translocated to the nucleus in murine B cells upon stimulation with CD40, whereas it was preferentially localized within the cytosol after stimulation with IgM, suggesting that signaling through CD40 and IgM differentially regulates ERK subcellular localization. Costimulation with CD40 and IgM (CD40/IgM) resulted in subcellular localization of ERK within the cytosol, supporting the notion that stimulation with IgM delivers the signal responsible for inhibition of ERK nuclear transport. Consistent with these observations, IgM and CD40/IgM stimulation resulted in activation of ribosomal S6 kinase, which is a cytoplasmic substrate for ERK, whereas CD40 stimulation had little effect on its activity. Disruption of the microtubule by colchicine in WEHI231 cells resulted in reduction of ERK activity in IgM signaling, but not in CD40 signaling, compatible with the notion that the microtubule network may hold cytoplasmic ERK activity mediated by IgM stimulation. These results support the notion that ERK could mediate different effector functions in B cells upon stimulation with IgM and CD40.
Shirakata
Y
Y
Department of Immunology, National Institute of Infectious Diseases, Tokyo, Japan.
Ishii
K
K
Yagita
H
H
Okumura
K
K
Taniguchi
M
M
Takemori
T
T
D000818:Animals; D001402:B-Lymphocytes / Q000201:enzymology* / Q000276:immunology*; D019013:CD40 Antigens / Q000494:pharmacology* / Q000502:physiology; D002478:Cells, Cultured; D003078:Colchicine / Q000633:toxicity; D003593:Cytoplasm / Q000201:enzymology; D004789:Enzyme Activation / Q000187:drug effects / Q000276:immunology; D007075:Immunoglobulin M / Q000494:pharmacology* / Q000502:physiology; D051379:Mice; D008810:Mice, Inbred C57BL; D008870:Microtubules / Q000187:drug effects; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism*; D019893:Ribosomal Protein S6 Kinases / Q000378:metabolism; D015398:Signal Transduction / Q000276:immunology; D013347:Subcellular Fractions / Q000201:enzymology / Q000276:immunology; D013379:Substrate Specificity / Q000276:immunology
D019013:CD40 Antigens; D007075:Immunoglobulin M; D019893:Ribosomal Protein S6 Kinases; D020928:Mitogen-Activated Protein Kinases; D003078:Colchicine