Transport of protein kinase C alpha into the nucleus requires intact cytoskeleton while the transport of a protein containing a canonical nuclear localization signal does not.
Protein kinase C undergoes a redistribution from the cytosol into the nucleus upon various stimuli. Since protein kinase C does not contain any known nuclear localization signal, the exact pathway and mechanism of the translocation into the nucleus is not known. We used immunofluorescence microscopy to investigate the role of the cytoskeleton in this process, and to detect the subcellular distribution of protein kinase C alpha in NIH 3T3 fibroblasts. In these cells protein kinase C alpha is translocated into the nucleus after stimulation with phorbol ester. We observed that cells treated with the cytoskeleton disrupting agents cytochalasin B or colchicine do not show the nuclear translocation of protein kinase C alpha after stimulation. In contrast, the nuclear accumulation of a nuclear localization signal containing reporter protein in an in vitro nuclear transport assay is not affected by these drugs. This observation has been confirmed for intact cells by microinjection experiments: cells which have been incubated with cytochalasin B or colchicine prior to microinjection of the reporter protein show the same accumulation in the nucleus as untreated cells. Our data show that intact cytoskeleton plays an important role in the translocation of protein kinase C alpha into the nucleus but not in the nuclear import of a karyophilic reporter protein.
Schmalz
D
D
Arbeitsgruppe Neurochemie, Freie Universität Berlin.
Kalkbrenner
F
F
Hucho
F
F
Buchner
K
K
D016475:3T3 Cells; D000595:Amino Acid Sequence; D000818:Animals; D001693:Biological Transport, Active; D002467:Cell Nucleus / Q000201:enzymology* / Q000378:metabolism; D003599:Cytoskeleton / Q000201:enzymology* / Q000378:metabolism; D005456:Fluorescent Dyes; D007527:Isoenzymes / Q000378:metabolism*; D051379:Mice; D008856:Microscopy, Fluorescence; D019913:Nuclear Localization Signals; D009687:Nuclear Proteins / Q000737:chemistry / Q000378:metabolism; D011493:Protein Kinase C / Q000378:metabolism*; D051571:Protein Kinase C-alpha; D028901:alpha Karyopherins
D005456:Fluorescent Dyes; D007527:Isoenzymes; D019913:Nuclear Localization Signals; D009687:Nuclear Proteins; D028901:alpha Karyopherins; C496630:Prkca protein, mouse; D011493:Protein Kinase C; D051571:Protein Kinase C-alpha