CRM1-mediated recycling of snurportin 1 to the cytoplasm.
Importin beta is a major mediator of import into the cell nucleus. Importin beta binds cargo molecules either directly or via two types of adapter molecules, importin alpha, for import of proteins with a classical nuclear localization signal (NLS), or snurportin 1, for import of m3G-capped U snRNPs. Both adapters have an NH2-terminal importin beta-binding domain for binding to, and import by, importin beta, and both need to be returned to the cytoplasm after having delivered their cargoes to the nucleus. We have shown previously that CAS mediates export of importin alpha. Here we show that snurportin 1 is exported by CRM1, the receptor for leucine-rich nuclear export signals (NESs). However, the interaction of CRM1 with snurportin 1 differs from that with previously characterized NESs. First, CRM1 binds snurportin 1 50-fold stronger than the Rev protein and 5,000-fold stronger than the minimum Rev activation domain. Second, snurportin 1 interacts with CRM1 not through a short peptide but rather via a large domain that allows regulation of affinity. Strikingly, snurportin 1 has a low affinity for CRM1 when bound to its m3G-capped import substrate, and a high affinity when substrate-free. This mechanism appears crucial for productive import cycles as it can ensure that CRM1 only exports snurportin 1 that has already released its import substrate in the nucleus.
Paraskeva
E
E
Zentrum für Molekulare Biologie der Universität Heidelberg, D-69120 Heidelberg, Germany.
Izaurralde
E
E
Bischoff
F R
FR
Huber
J
J
Kutay
U
U
Hartmann
E
E
Lührmann
R
R
Görlich
D
D
D000818:Animals; D001665:Binding Sites; D002352:Carrier Proteins / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism; D002846:Chromatography, Affinity; D003593:Cytoplasm / Q000378:metabolism; D004926:Escherichia coli; D005260:Female; D006367:HeLa Cells; D006801:Humans; D028884:Karyopherins; D007700:Kinetics; D008954:Models, Biological; D009687:Nuclear Proteins / Q000302:isolation & purification / Q000378:metabolism; D009865:Oocytes / Q000502:physiology; D039381:RNA Cap-Binding Proteins; D018160:Receptors, Cytoplasmic and Nuclear / Q000302:isolation & purification / Q000378:metabolism*; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D017411:Ribonucleoproteins, Small Nuclear / Q000378:metabolism*; D015398:Signal Transduction; D014157:Transcription Factors / Q000378:metabolism*; D014982:Xenopus laevis; D000097606:Exportin 1 Protein
D002352:Carrier Proteins; D028884:Karyopherins; D009687:Nuclear Proteins; D039381:RNA Cap-Binding Proteins; D018160:Receptors, Cytoplasmic and Nuclear; D011994:Recombinant Proteins; D017411:Ribonucleoproteins, Small Nuclear; D014157:Transcription Factors; D000097606:Exportin 1 Protein; C114090:SNUPN protein, human