Importin beta-depending nuclear import pathways: role of the adapter proteins in the docking and releasing steps.
Nuclear imports of uridine-rich small nuclear ribonucleoprotein (U1 snRNP) and proteins with classical nuclear localization signal (cNLS-protein) are mediated by importin beta. However, due to the presence of different import signals, the adapter protein of the imported molecules and importin beta is different for each pathway. Although the adapter for cNLS-protein is importin alpha, the adapter for U1 snRNP is snurportin1 (SPN1). Herein, we show that the use of distinct adapters by importin beta results in differences at the docking and releasing step for these two import pathways. Nuclear pore complex (NPC) docking of U1 snRNP but not of cNLS-protein was inhibited by an anti-CAN/Nup214 antibody. Thus, the initial NPC-binding site is different for each pathway. Pull-down assays between immobilized SPN1 and two truncated forms of importin beta documented that SPN1 and importin alpha have different binding sites on importin beta. Importin beta fragment 1-618, which binds to SPN1 but not to importin alpha, was able to support the nuclear import of U1 snRNPs. After the translocation through the NPC, both import complexes associated with the nuclear side of the NPC. However, we found that the nature of the importin beta-binding domain of the adapters influences the release of the cargo into the nucleoplasm.
Rollenhagen
Christiane
C
Department of Zoology, University of British Columbia, Vancouver, V6T 1Z4 Canada.
Mühlhäusser
Petra
P
Kutay
Ulrike
U
Panté
Nelly
N
D021581:Active Transport, Cell Nucleus / Q000502:physiology; D033942:Adaptor Proteins, Vesicular Transport / Q000378:metabolism*; D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism / Q000648:ultrastructure; D028861:Nuclear Pore Complex Proteins / Q000378:metabolism; D009865:Oocytes / Q000378:metabolism / Q000648:ultrastructure; D017434:Protein Structure, Tertiary; D021381:Protein Transport / Q000502:physiology; D017411:Ribonucleoproteins, Small Nuclear / Q000378:metabolism; D014981:Xenopus; D028901:alpha Karyopherins / Q000378:metabolism; D028961:beta Karyopherins / Q000378:metabolism*
D033942:Adaptor Proteins, Vesicular Transport; D028861:Nuclear Pore Complex Proteins; D017411:Ribonucleoproteins, Small Nuclear; D028901:alpha Karyopherins; D028961:beta Karyopherins