Nuclear shuttling of mitogen-activated protein (MAP) kinase (extracellular signal-regulated kinase (ERK) 2) was dynamically controlled by MAP/ERK kinase after antigen stimulation in RBL-2H3 cells.
The mitogen-activated protein kinase (MAPK) cascade consists of the MAPK (extracellular signal-regulated kinase 2; ERK2) and its activator, MAPK kinase (MAP/ERK kinase; MEK). However, the mechanisms for activation of ERK2 have not been defined yet in cells. Here, we used fluorescent protein-tagged ERK2 and MEK to examine the localization of ERK2 and MEK in living rat basophilic leukemia (RBL-2H3) cells. ERK2 was mainly in the cytoplasm in resting cells but translocated into the nucleus after the ligation of IgE receptors. The import of ERK2 reached the maximum at 6--7 min, and then the imported ERK2 was exported from the nucleus. MEK mainly resided in the cytoplasm, and no significant MEK translocation was detected statically after ligation of IgE receptors. However, analysis of the dynamics of ERK2 and MEK suggested that both of them rapidly shuttle between the cytoplasm and the nucleus and that MEK regulates the nuclear shuttling of ERK2, whereas MEK remains mainly in the cytoplasm. In addition, the data suggested that the sustained calcium increase was required for the optimal translocation of ERK2 into the nucleus in RBL-2H3 cells. These results gave a new insight of the dynamics of ERK2 and MEK in the nuclear shuttling of RBL-2H3 cells after the ligation of IgE receptors.
Faculty of Pharmaceutical Sciences, Nagoya City University, Nagoya, Japan.
T
T
Furuno
N
N
Hirashima
S
S
Onizawa
N
N
Sagiya
M
M
Nakanishi
D021581:Active Transport, Cell Nucleus / Q000235:genetics / Q000276:immunology; D000818:Animals; D000941:Antigens / Q000276:immunology*; D001426:Bacterial Proteins / Q000235:genetics; D002118:Calcium / Q000502:physiology; D002413:Cations, Divalent / Q000494:pharmacology; D002467:Cell Nucleus / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D004140:Dinitrophenols / Q000276:immunology; D004789:Enzyme Activation / Q000235:genetics; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D019950:Mitogen-Activated Protein Kinase 1 / Q000235:genetics / Q000378:metabolism*; D020929:Mitogen-Activated Protein Kinase Kinases / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D012710:Serum Albumin, Bovine / Q000276:immunology; D013347:Subcellular Fractions / Q000201:enzymology; D014162:Transfection; D014407:Tumor Cells, Cultured / Q000201:enzymology* / Q000378:metabolism
D000941:Antigens; D001426:Bacterial Proteins; D002413:Cations, Divalent; D004140:Dinitrophenols; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; C034638:dinitrophenyl-bovine serum albumin; C054562:yellow fluorescent protein, Bacteria; D049452:Green Fluorescent Proteins; D012710:Serum Albumin, Bovine; D019950:Mitogen-Activated Protein Kinase 1; D020929:Mitogen-Activated Protein Kinase Kinases; D002118:Calcium