title: Cell cycle-dependent phosphorylation of nucleoporins and nuclear pore membrane protein Gp210.
abstract: During mitosis in higher eukaryotic cells, the nuclear envelope membranes break down into distinct populations of vesicles and the proteins of the nuclear lamina and the nuclear pore complexes disperse in the cytoplasm. Since phosphorylation can alter protein-protein interactions and membrane traffic, we have examined the cell cycle-dependent phosphorylation of nuclear pore complex proteins. Nonmembrane nucleoporins Nup153, Nup214, and Nup358 that are modified by O-linked N-acetylglucosamine and recognized by a monoclonal antibody were phosphorylated throughout the cell cycle and hyperphosphorylated during M phase. Pore membrane glycoprotein gp210, that has a cytoplasmic, carboxyl-terminal domain facing the pore, was not phosphorylated in interphase but specifically phosphorylated in mitosis. Mutant and wild-type fusion proteins containing the cytoplasmic domain of gp210 were phosphorylated in vitro and their phosphopeptide maps compared to that of mitotic gp210. This analysis showed that Ser1880 of gp210 was phosphorylated in mitosis, possibly by cyclin B-p34cdc2 or a related kinase. Several nuclear pore complex proteins are therefore differentially phosphorylated during mitosis when pore complexes disassemble and reassemble.
authors.affiliation: Département de Biologie Cellulaire, Institut Jacques Monod, CNRS, Université Paris 7, France.
authors.forename: C
authors.initials: C
authors.lastname: Favreau
authors.forename: H J
authors.initials: HJ
authors.lastname: Worman
authors.forename: R W
authors.initials: RW
authors.lastname: Wozniak
authors.forename: T
authors.initials: T
authors.lastname: Frappier
authors.forename: J C
authors.initials: JC
authors.lastname: Courvalin
mesh_terms: D000117:Acetylglucosamine; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D001323:Autoantibodies; D001483:Base Sequence; D002453:Cell Cycle; D002460:Cell Line; D017931:DNA Primers; D006367:HeLa Cells; D006801:Humans; D008114:Liver Neoplasms, Experimental; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009685:Nuclear Envelope / Q000378:metabolism / Q000648:ultrastructure; D028861:Nuclear Pore Complex Proteins; D009687:Nuclear Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D009838:Oligodeoxyribonucleotides; D010766:Phosphorylation; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D012694:Serine
chemical_list: D000911:Antibodies, Monoclonal; D001323:Autoantibodies; D017931:DNA Primers; D008562:Membrane Glycoproteins; C063865:NUP210 protein, human; D028861:Nuclear Pore Complex Proteins; D009687:Nuclear Proteins; D009838:Oligodeoxyribonucleotides; D011993:Recombinant Fusion Proteins; D012694:Serine; D000117:Acetylglucosamine