title: The RNA binding activity of a ribosome biogenesis factor, nucleophosmin/B23, is modulated by phosphorylation with a cell cycle-dependent kinase and by association with its subtype.
abstract: Nucleophosmin/B23 is a nucleolar phosphoprotein. It has been shown that B23 binds to nucleic acids, digests RNA, and is localized in nucleolar granular components from which preribosomal particles are transported to cytoplasm. The intracellular localization of B23 is significantly changed during the cell cycle. Here, we have examined the cellular localization of B23 proteins and the effect of mitotic phosphorylation of B23.1 on its RNA binding activity. Two splicing variants of B23 proteins, termed B23.1 and B23.2, were complexed both in vivo and in vitro. The RNA binding activity of B23.1 was impaired by hetero-oligomer formation with B23.2. Both subtypes of B23 proteins were phosphorylated during mitosis by cyclin B/cdc2. The RNA binding activity of B23.1 was repressed through cyclin B/cdc2-mediated phosphorylation at specific sites in B23. Thus, the RNA binding activity of B23.1 is stringently modulated by its phosphorylation and subtype association. Interphase B23.1 was mainly localized in nucleoli, whereas B23.2 and mitotic B23.1, those of which were incapable of binding to RNA, were dispersed throughout the nucleoplasm and cytoplasm, respectively. These results suggest that nucleolar localization of B23.1 is mediated by its ability to associate with RNA.
authors.affiliation: Department of Infection Biology, Institute of Basic Medical Sciences, University of Tsukuba, 1-1-1 Tennohdai, Tsukuba 305-8575, Japan.
authors.forename: Mitsuru
authors.initials: M
authors.lastname: Okuwaki
authors.forename: Masafumi
authors.initials: M
authors.lastname: Tsujimoto
authors.forename: Kyosuke
authors.initials: K
authors.lastname: Nagata
mesh_terms: D000199:Actins / Q000235:genetics; D017398:Alternative Splicing; D019943:Amino Acid Substitution; D000818:Animals; D001483:Base Sequence; D002466:Cell Nucleolus / Q000502:physiology; D002645:Chickens; D018844:Cyclin-Dependent Kinases / Q000378:metabolism*; D017931:DNA Primers; D003850:Deoxyribonuclease I; D004742:Enhancer Elements, Genetic; D017781:Genes, Immediate-Early; D014644:Genetic Variation; D006367:HeLa Cells; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism; D016297:Mutagenesis, Site-Directed; D009687:Nuclear Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000090243:Nucleophosmin; D010766:Phosphorylation; D011401:Promoter Regions, Genetic; D020033:Protein Isoforms / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012259:Ribonuclease, Pancreatic; D012270:Ribosomes / Q000502:physiology* / Q000648:ultrastructure
chemical_list: D000199:Actins; D017931:DNA Primers; D007527:Isoenzymes; C000716967:NPM1 protein, human; D009687:Nuclear Proteins; D020033:Protein Isoforms; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins; D000090243:Nucleophosmin; D018844:Cyclin-Dependent Kinases; D003850:Deoxyribonuclease I; D012259:Ribonuclease, Pancreatic