Abstract
Nuclear import of the four core histones H2A, H2B, H3 and H4 is one of the main nuclear import activities during S-phase of the cell cycle. However, the molecular machinery facilitating nuclear import of core histones has not been elucidated. Here, we investigated the pathways by which histone import can occur. First, we show that core histone import can be competed by the BIB (beta-like import receptor binding) domain of ribosomal protein L23a suggesting that histone import is an importin mediated process. Secondly, affinity chromatography on immobilized core histones revealed that several members of the importin beta family of transport receptors are able to interact with core histones. Finally, we demonstrate that at least four known and one novel importin, importin 9, can mediate nuclear import of core histones into the nuclei of permeabilized cells. Our results suggest that multiple pathways of import exist to provide efficient nuclear uptake of these abundant, essential proteins.
MeSH Terms
Active Transport, Cell Nucleus/physiology
Animals
Cell Nucleus/metabolism
Cloning, Molecular
Genes, Reporter
HeLa Cells
Histones/metabolism
Humans
Karyopherins/genetics,metabolism
Microscopy, Confocal
Molecular Sequence Data
Peptide Fragments/metabolism
Protein Binding
Rabbits
Recombinant Fusion Proteins/genetics,metabolism
S Phase/physiology
ran GTP-Binding Protein/metabolism
Chemicals
Histones
Karyopherins
Peptide Fragments
Recombinant Fusion Proteins
ran GTP-Binding Protein
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Mühlhäusser P
ETH Zürich, Institut für Biochemie, Universitätsstrasse 16, 8092 Zürich, Switzerland and 1Max-Delbrück-Centrum, Robert-Rössle-Strasse 10, 13122 Berlin-Buch, Germany.
Müller E C
Otto A
Kutay U
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