Characterization of an unusual importin alpha binding motif in the borna disease virus p10 protein that directs nuclear import.
Nuclear import of many cellular and viral proteins is mediated by short nuclear localization signals (NLS) that are recognized by intracellular receptor proteins belonging to the importin/karyopherin alpha and beta families. The primary structure of NLS is not well defined, but most contain at least three basic amino acids and harbor the relative consensus sequence K(K/R)X(K/R). We have studied the nuclear import of the Borna disease virus p10 protein that lacks a canonical oligobasic NLS. It is shown that the p10 protein exhibits all characteristics of an actively transported molecule in digitonin-permeabilized cells. Import activity was found to reside in the 20 N-terminal p10 amino acids that are devoid of an NLS consensus motif. Unexpectedly, p10-dependent import was blocked by a peptide inhibitor of importin alpha-dependent nuclear translocation, and the transport activity of the p10 N-terminal domain was shown to correlate with the ability to bind to importin alpha. These findings suggest that nuclear import of the Borna disease virus p10 protein occurs through a nonconventional karyophilic signal and highlight that the cellular importin alpha NLS receptor proteins can recognize nuclear targeting signals that substantially deviate from the consensus sequence.
Wolff
Thorsten
T
Robert-Koch-Institut, Nordufer 20, 13353 Berlin, Germany. wolfft@rki.de
Unterstab
Gunhild
G
Heins
Gudrun
G
Richt
Juergen A
JA
Kann
Michael
M
D021581:Active Transport, Cell Nucleus; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism*; D004252:DNA Mutational Analysis; D005982:Glutathione Transferase / Q000378:metabolism; D006801:Humans; D018613:Microscopy, Confocal; D008969:Molecular Sequence Data; D019913:Nuclear Localization Signals; D011233:Precipitin Tests; D011485:Protein Binding; D020033:Protein Isoforms; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D014764:Viral Proteins / Q000378:metabolism*; D028901:alpha Karyopherins / Q000737:chemistry* / Q000378:metabolism*
D019913:Nuclear Localization Signals; D020033:Protein Isoforms; D014764:Viral Proteins; D028901:alpha Karyopherins; C119397:p10 protein, Borna disease virus; D005982:Glutathione Transferase