Importin 13 mediates nuclear import of histone fold-containing chromatin accessibility complex heterodimers.
The histone fold is a structural element that facilitates heterodimerization, and histone fold heterodimers play crucial roles in gene regulation. Here, we investigated the nuclear import of two human histone fold pairs, which belong to the H2A/H2B family: CHRAC-15/CHRAC-17 and p12/CHRAC-17. Our results from in vitro nuclear import assays with permeabilized cells and in vivo cotransfection experiments reveal that importin 13 facilitates nuclear import of both histone fold heterodimers. Using glutathione S-transferase pulldown experiments, we provide evidence that heterodimers are required for efficient binding of importin 13 because the monomers alone do not significantly interact. Mutational analysis shows that stepwise substitution of basic amino acid residues conserved among the histone fold subunits leads to a progressive loss of importin 13 binding and nuclear accumulation of CHRAC-15/CHRAC-17 and p12/CHRAC-17. The distribution of basic amino acid residues among the histone fold subunits essential for nuclear uptake suggests that heterodimerization of the histone fold motif-containing proteins forms an importin 13-specific binding platform.
Walker
Patrick
P
Institut für Biochemie und Molekulare Zellbiologie, Abteilung Molekularbiologie, Universität Göttingen, Humboldtallee 23, 37073 Göttingen, Germany.
Doenecke
Detlef
D
Kahle
Joerg
J
D021581:Active Transport, Cell Nucleus; D020816:Amino Acid Motifs; D002843:Chromatin / Q000737:chemistry; D003593:Cytoplasm / Q000378:metabolism; D004252:DNA Mutational Analysis; D019281:Dimerization; D005982:Glutathione Transferase / Q000378:metabolism; D006367:HeLa Cells; D006657:Histones / Q000737:chemistry; D006801:Humans; D028884:Karyopherins / Q000378:metabolism*; D008954:Models, Biological; D016297:Mutagenesis, Site-Directed; D017510:Protein Folding; D011994:Recombinant Proteins / Q000737:chemistry
D002843:Chromatin; D006657:Histones; C527880:IPO13 protein, human; D028884:Karyopherins; D011994:Recombinant Proteins; D005982:Glutathione Transferase