Catalytic activity of protein kinase CK1 delta (casein kinase 1delta) is essential for its normal subcellular localization.
Mammalian casein kinase 1delta (CK1delta) is a homologue of the S. cerevisiae Hrr25p protein kinase. Hrr25p is involved in regulating diverse events including vesicular trafficking, gene expression, DNA repair, and chromosome segregation. In contrast to Hrr25p, little is known about the function, regulation, or subcellular localization of CK1delta. In the present study, we show that CK1delta in mammalian cells is mainly cytoplasmic and enriched within the Golgi and/or ER-Golgi transport vesicles, consistent with a role in vesicular trafficking. Transient expression of green fluorescent protein (GFP)- or FLAG peptide-tagged CK1delta showed localization similar to that of the endogenous CK1delta. GFP-CK1delta was also enriched at the centrosomes in interphase cells. Strikingly, two inactive mutant CK1delta proteins (K38M and T176I) showed almost exclusive nuclear staining, suggesting that protein kinase activity is required for normal localization of CK1delta and prevention of nuclear accumulation. The nuclear export inhibitor leptomycin B promoted nuclear enrichment of CK1delta indicating that nuclear localization of CK1delta occurs physiologically. Both endogenous CK1delta and GFP-CK1delta are enriched on the spindle poles in mitotic cells, consistent with a role in regulating spindle formation. Localization is a property of the protein kinase domain and is independent of the C-terminal noncatalytic domain. These data are consistent with roles for CK1delta in mammalian cells analogous to those of its yeast counterparts.
Milne
D M
DM
Biomedical Research Centre, Ninewells Hospital and Medical School, Dundee, DD1 9SY, United Kingdom.
Looby
P
P
Meek
D W
DW
D000818:Animals; D047388:Casein Kinases; D020134:Catalytic Domain; D002453:Cell Cycle / Q000502:physiology; D002460:Cell Line; D002467:Cell Nucleus / Q000201:enzymology*; D018385:Centrosome / Q000201:enzymology; D003593:Cytoplasm / Q000201:enzymology; D004721:Endoplasmic Reticulum / Q000201:enzymology; D005231:Fatty Acids, Unsaturated / Q000494:pharmacology; D017930:Genes, Reporter; D006056:Golgi Apparatus / Q000201:enzymology / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D007399:Interphase; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D008938:Mitosis; D011494:Protein Kinases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D008941:Spindle Apparatus / Q000201:enzymology; D014162:Transfection; D022161:Transport Vesicles / Q000201:enzymology
D005231:Fatty Acids, Unsaturated; D007527:Isoenzymes; D011993:Recombinant Fusion Proteins; D011494:Protein Kinases; D047388:Casein Kinases; C038753:leptomycin B