The Journal of cell biology, 2011-08, Vol.194 (4), p.597-612
2011
Details
- Autor(en) / Beteiligte
- Titel
- Ran-dependent docking of importin-beta to RanBP2/Nup358 filaments is essential for protein import and cell viability
- Ist Teil von
- The Journal of cell biology, 2011-08, Vol.194 (4), p.597-612
- Ort / Verlag
- United States: The Rockefeller University Press
- Erscheinungsjahr
- 2011
- Link zum Volltext
- Quelle
- EZB Free E-Journals
- Beschreibungen/Notizen
- RanBP2/Nup358, the major component of the cytoplasmic filaments of the nuclear pore complex (NPC), is essential for mouse embryogenesis and is implicated in both macromolecular transport and mitosis, but its specific molecular functions are unknown. Using RanBP2 conditional knockout mouse embryonic fibroblasts and a series of mutant constructs, we show that transport, rather than mitotic, functions of RanBP2 are required for cell viability. Cre-mediated RanBP2 inactivation caused cell death with defects in M9- and classical nuclear localization signal (cNLS)-mediated protein import, nuclear export signal-mediated protein export, and messenger ribonucleic acid export but no apparent mitotic failure. A short N-terminal RanBP2 fragment harboring the NPC-binding domain, three phenylalanine-glycine motifs, and one Ran-binding domain (RBD) corrected all transport defects and restored viability. Mutation of the RBD within this fragment caused lethality and perturbed binding to Ran guanosine triphosphate (GTP)-importin-β, accumulation of importin-β at nuclear pores, and cNLS-mediated protein import. These data suggest that a critical function of RanBP2 is to capture recycling RanGTP-importin-β complexes at cytoplasmic fibrils to allow for adequate cNLS-mediated cargo import.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 0021-9525eISSN: 1540-8140DOI: 10.1083/jcb.201102018Titel-ID: cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_3160583
- Format
- –
- Schlagworte
- Active Transport, Cell Nucleus, Animals, beta Karyopherins - genetics, beta Karyopherins - metabolism, Binding Sites, Cell Line, Cell Proliferation, Cell Survival, Chromosome Segregation, Cytoskeleton - metabolism, Fibroblasts - metabolism, Heterogeneous-Nuclear Ribonucleoprotein Group A-B - metabolism, Humans, Mice, Mice, Knockout, Microscopy, Fluorescence, Microscopy, Video, Mitosis, Molecular Chaperones - genetics, Molecular Chaperones - metabolism, Mutation, Nuclear Localization Signals - metabolism, Nuclear Pore - metabolism, Nuclear Pore Complex Proteins - deficiency, Nuclear Pore Complex Proteins - genetics, Nuclear Pore Complex Proteins - metabolism, Peptide Fragments - metabolism, Protein Interaction Domains and Motifs, Protein Interaction Mapping, Protein Sorting Signals, ran GTP-Binding Protein - genetics, ran GTP-Binding Protein - metabolism, Recombinant Fusion Proteins - metabolism, RNA, Messenger - metabolism, Small Ubiquitin-Related Modifier Proteins - metabolism, Time Factors, Transfection, Ubiquitin-Protein Ligases - metabolism