title: Phosphorylation on protein kinase C sites inhibits nuclear import of lamin B2.
abstract: The nuclear lamina is a karyoskeletal structure at the nucleoplasmic surface of the inner nuclear membrane. Its assembly state is regulated by phosphorylation of the intermediate filament type lamin proteins. Strong evidence has been obtained for a causal link between phosphorylation of lamins by the p34cdc2 protein kinase and disassembly of the nuclear lamina during mitosis. In contrast, no information is currently available on the role of lamin phosphorylation during interphase of the cell cycle. Here, we have identified four protein kinase C phosphorylation sites in purified chicken lamin B2 as serines 400, 404, 410, and 411. In vivo, the tryptic peptide containing serines 400 and 404 is phosphorylated throughout interphase, whereas serines 410 and 411 become phosphorylated specifically in response to activation of protein kinase C by phorbol ester. Prompted by the close proximity of serines 410/411 to the nuclear localization signal of lamin B2, we have studied the influence of phosphorylation of these residues on nuclear transport. Using an in vitro assay, we show that phosphorylation of lamin B2 by protein kinase C strongly inhibits transport to the nucleus. Moreover, phorbol ester treatment of intact cells leads to a substantial reduction of the rate of nuclear import of newly synthesized lamin B2 in vivo. These findings have implications for the dynamic structure of the nuclear lamina, and they suggest that the modulation of nuclear transport rates by cytoplasmic phosphorylation may represent a general mechanism for regulating nuclear activities.
authors.affiliation: Swiss Institute for Experimental Cancer Research (ISREC), Epalinges.
authors.forename: H
authors.initials: H
authors.lastname: Hennekes
authors.forename: M
authors.initials: M
authors.lastname: Peter
authors.forename: K
authors.initials: K
authors.lastname: Weber
authors.forename: E A
authors.initials: EA
authors.lastname: Nigg
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism*; D002645:Chickens; D006367:HeLa Cells; D006801:Humans; D007399:Interphase; D034921:Lamin Type B; D034882:Lamins; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D010449:Peptide Mapping; D010748:Phosphopeptides / Q000302:isolation & purification; D010766:Phosphorylation; D011493:Protein Kinase C / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D017384:Sequence Deletion; D012694:Serine; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014407:Tumor Cells, Cultured
chemical_list: D034921:Lamin Type B; D034882:Lamins; D046911:Macromolecular Substances; D009687:Nuclear Proteins; D010748:Phosphopeptides; D011994:Recombinant Proteins; D012694:Serine; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate