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Nxt1 Is Necessary for the Terminal Step of Crm1-Mediated Nuclear Export
- Source :
- The Journal of Cell Biology, Journal of Cell Biology, Journal of Cell Biology, Rockefeller University Press, 2001, 152 (1), pp.141-156. ⟨10.1083/jcb.152.1.141⟩
- Publication Year :
- 2001
- Publisher :
- The Rockefeller University Press, 2001.
-
Abstract
- Soluble factors are required to mediate nuclear export of protein and RNA through the nuclear pore complex (NPC). These soluble factors include receptors that bind directly to the transport substrate and regulators that determine the assembly state of receptor–substrate complexes. We recently reported the identification of NXT1, an NTF2-related export factor that stimulates nuclear protein export in permeabilized cells and undergoes nucleocytoplasmic shuttling in vivo (Black, B.E., L. Lévesque, J.M. Holaska, T.C. Wood, and B.M. Paschal. 1999. Mol. Cell. Biol. 19:8616–8624). Here, we describe the molecular characterization of NXT1 in the context of the Crm1-dependent export pathway. We find that NXT1 binds directly to Crm1, and that the interaction is sensitive to the presence of Ran-GTP. Moreover, mutations in NXT1 that reduce binding to Crm1 inhibit the activity of NXT1 in nuclear export assays. We show that recombinant Crm1 and Ran are sufficient to reconstitute nuclear translocation of a Rev reporter protein from the nucleolus to an antibody accessible site on the cytoplasmic side of the NPC. Further progress on the export pathway, including the terminal step of Crm1 and Rev reporter protein release, requires NXT1. We propose that NXT1 engages with the export complex in the nucleoplasm, and that it facilitates delivery of the export complex to a site on the cytoplasmic side of NPC where the receptor and substrate are released into the cytoplasm.
- Subjects :
- green fluorescent protein
Nucleocytoplasmic Transport Proteins
Time Factors
[SDV]Life Sciences [q-bio]
NXT1
STV
Receptors, Cytoplasmic and Nuclear
Crm1
0302 clinical medicine
RNA, Transfer
glutathione-S -transferase
Genes, Reporter
RNA, Small Nuclear
glucocorticoid receptor
Nuclear protein
Nuclear pore
leptomycin B
0303 health sciences
PKI
protein kinase inhibitor
nuclear export signal
nuclear envelope
Cell biology
GR
medicine.anatomical_structure
Gene Products, rev
Original Article
NPC
NE
Recombinant Fusion Proteins
Active Transport, Cell Nucleus
NLS
nuclear transport
Biology
Karyopherins
GFP
Cell Line
03 medical and health sciences
Rev response element
nuclear pore complex
constitutive transport element
medicine
streptavidin
Animals
RNA, Messenger
GST
Nuclear export signal
LMB
030304 developmental biology
Cell Nucleus
Nucleoplasm
Cell Biology
nuclear localization signal
CTE
Cell nucleus
Ran
Cytoplasm
Mutagenesis
NES
RRE
Carrier Proteins
030217 neurology & neurosurgery
Subjects
Details
- Language :
- English
- ISSN :
- 15408140 and 00219525
- Volume :
- 152
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- The Journal of Cell Biology
- Accession number :
- edsair.doi.dedup.....3cfe7dddd172b452cedb5ea357b9c336