Importin beta contains a COOH-terminal nucleoporin binding region important for nuclear transport.
Proteins containing a classical NLS are transported into the nucleus by the import receptor importin beta, which binds to cargoes via the adaptor importin alpha. The import complex is translocated through the nuclear pore complex by interactions of importin beta with a series of nucleoporins. Previous studies have defined a nucleoporin binding region in the NH2-terminal half of importin beta. Here we report the identification of a second nucleoporin binding region in its COOH-terminal half. Although the affinity of the COOH-terminal region for nucleoporins is dramatically weaker than that of the NH2-terminal region, sets of mutations that perturb the nucleoporin binding of either region reduce the nuclear import activity of importin beta to a similar extent ( approximately 50%). An importin beta mutant with a combination of mutations in the NH2- and COOH-terminal regions is completely inactive for nuclear import. Thus, importin beta possesses two nucleoporin binding sites, both of which are important for its nuclear import function.
Bednenko
Janna
J
Department of Cell Biology, The Scripps Research Institute, La Jolla, CA 92037, USA.
Cingolani
Gino
G
Gerace
Larry
L
D021581:Active Transport, Cell Nucleus / Q000235:genetics / Q000502:physiology*; D000596:Amino Acids / Q000378:metabolism; D001665:Binding Sites / Q000235:genetics; D002264:Carboxylic Acids / Q000378:metabolism; D015536:Down-Regulation / Q000235:genetics; D005057:Eukaryotic Cells / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D022022:Nuclear Pore / Q000235:genetics / Q000378:metabolism*; D028861:Nuclear Pore Complex Proteins / Q000378:metabolism*; D017354:Point Mutation / Q000235:genetics; D017434:Protein Structure, Tertiary / Q000235:genetics; D021381:Protein Transport / Q000235:genetics; D028961:beta Karyopherins / Q000235:genetics / Q000378:metabolism*
D000596:Amino Acids; D002264:Carboxylic Acids; D028861:Nuclear Pore Complex Proteins; D028961:beta Karyopherins