title: The requirement of H1 histones for a heterodimeric nuclear import receptor.
abstract: After synthesis in the cytoplasm, H1 histones are imported into the nucleus through an energy-dependent process that can be mediated by an importin beta-importin 7 (Impbeta-Imp7) heterodimer. H1 histones contain two structurally different types of nuclear localization signals (NLS). The first type of NLS resides within the unstructured C-terminal domain and is rich in basic amino acids. In contrast, the highly conserved central domain of the H1 histone contains comparatively few basic amino acids but also represents a functional NLS. The competence for the nuclear import of this globular domain seems to be based on its secondary structure. Here, we show that the Impbeta-Imp7 heterodimer is the only receptor for H1 import. Furthermore, we identified the import receptors mediating the in vitro transport of different NLS of the H1 histone. Using the digitonin-permeabilized cell import assay we show that Impbeta is the most efficient import receptor for the globular domain of H1 histones, whereas the heterodimer of Impbeta and Imp7 is the functional receptor for the entire C-terminal domain. However, short fragments of the C-terminal domain are imported in vitro by at least four different importins, which resembles the import pathway of ribosomal proteins and core histones. In addition, we show that heterodimerization of Impbeta with Imp7 is absolutely necessary for their proper function as an import receptor for H1 histones. These findings point to a chaperone-like function of the heterodimeric complex in addition to its function as an import receptor. It appears that the Impbeta-Imp7 heterodimer is specialized for NLS consisting of extended basic domains.
authors.affiliation: Institut für Biochemie und Molekulare Zellbiologie, Abteilung für Molekularbiologie, Universität Göttingen, D-37073 Göttingen, Germany.
authors.forename: Marc
authors.initials: M
authors.lastname: Bäuerle
authors.forename: Detlef
authors.initials: D
authors.lastname: Doenecke
authors.forename: Werner
authors.initials: W
authors.lastname: Albig
mesh_terms: D021581:Active Transport, Cell Nucleus; D000818:Animals; D001665:Binding Sites; D002467:Cell Nucleus / Q000378:metabolism*; D017124:Conserved Sequence; D004072:Digitonin / Q000494:pharmacology; D019281:Dimerization; D004591:Electrophoresis, Polyacrylamide Gel; D005456:Fluorescent Dyes / Q000494:pharmacology; D017353:Gene Deletion; D006367:HeLa Cells; D006657:Histones / Q000737:chemistry / Q000378:metabolism* / Q000502:physiology; D006801:Humans; D028884:Karyopherins / Q000737:chemistry / Q000502:physiology*; D019913:Nuclear Localization Signals; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011817:Rabbits; D018160:Receptors, Cytoplasmic and Nuclear; D011994:Recombinant Proteins / Q000378:metabolism; D012156:Reticulocytes / Q000378:metabolism; D014162:Transfection; D028961:beta Karyopherins / Q000737:chemistry / Q000502:physiology*
chemical_list: D005456:Fluorescent Dyes; D006657:Histones; C466996:IPO7 protein, human; D028884:Karyopherins; D019913:Nuclear Localization Signals; D018160:Receptors, Cytoplasmic and Nuclear; D011994:Recombinant Proteins; D028961:beta Karyopherins; D004072:Digitonin