Disruption of the FG nucleoporin NUP98 causes selective changes in nuclear pore complex stoichiometry and function.
The NUP98 gene encodes precursor proteins that generate two nucleoplasmically oriented nucleoporins, NUP98 and NUP96. By using gene targeting, we have selectively disrupted the murine NUP98 protein, leaving intact the expression and localization of NUP96. We show that NUP98 is essential for mouse gastrulation, a developmental stage that is associated with rapid cell proliferation, but dispensable for basal cell growth. NUP98-/- cells had an intact nuclear envelope with a normal number of embedded nuclear pore complexes. Typically, NUP98-deficient cells contained on average approximately 5-fold more cytoplasmic annulate lamellae than control cells. We found that a set of cytoplasmically oriented nucleoporins, including NUP358, NUP214, NUP88, and p62, assembled inefficiently into nuclear pores of NUP98-/- cells. Instead, these nucleoporins were prominently associated with the annulate lamellae. By contrast, a group of nucleoplasmically oriented nucleoporins, including NUP153, NUP50, NUP96, and NUP93, had no affinity for annulate lamellae and assembled normally into nuclear pores. Mutant pores were significantly impaired in transport receptor-mediated docking of proteins with a nuclear localization signal or M9 import signal and showed weak nuclear import of such substrates. In contrast, the ability of mutant pores to import ribosomal protein L23a and spliceosome protein U1A appeared intact. These observations show that NUP98 disruption selectively impairs discrete protein import pathways and support the idea that transport of distinct import complexes through the nuclear pore complex is mediated by specific subsets of nucleoporins.
Wu
X
X
Department of Pediatrics and Adolescent Medicine, Mayo Clinic, Rochester, MN 55905, USA.
Kasper
L H
LH
Mantcheva
R T
RT
Mantchev
G T
GT
Springett
M J
MJ
van Deursen
J M
JM
D021581:Active Transport, Cell Nucleus; D000818:Animals; D001692:Biological Transport; D002455:Cell Division; D002467:Cell Nucleus / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism; D005775:Gastrula / Q000502:physiology; D018390:Gene Targeting; D006801:Humans; D028884:Karyopherins / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D016296:Mutagenesis; D022022:Nuclear Pore / Q000378:metabolism*; D028861:Nuclear Pore Complex Proteins / Q000235:genetics / Q000378:metabolism*; D018160:Receptors, Cytoplasmic and Nuclear; D000097606:Exportin 1 Protein
D028884:Karyopherins; D028861:Nuclear Pore Complex Proteins; D018160:Receptors, Cytoplasmic and Nuclear; D000097606:Exportin 1 Protein; C119239:nuclear pore complex protein 96; C093350:nuclear pore complex protein 98