G2 arrest and impaired nucleocytoplasmic transport in mouse embryos lacking the proto-oncogene CAN/Nup214.
The vertebrate nucleopore complex (NPC) is a 125 MDa multiprotein assembly that mediates nucleocytoplasmic transport. One of its components, CAN/Nup214, is an FXFG repeat-containing protein known to be involved in myeloid leukemia in humans. We have devised a powerful genetic approach, using maternally derived protein in murine null embryos, to show that CAN/ Nup214 is essential for NPC function in vivo. We demonstrate that CAN-/- mouse embryonic stem (ES) cells are not viable and that CAN-/- embryos die in utero between 4.0 and 4.5 days postcoitum, following the depletion of their CAN from maternal sources. In 3.5-day-old mutant embryos, cultured in vitro, progressive depletion of CAN leads to cell cycle arrest in G2 phase, and eventually to blastocoel collapse, impaired NLS-mediated protein uptake and nuclear accumulation of polyadenylated RNA. Remarkably, these defective CAN-depleted embryos do not display any gross morphological abnormalities in their nuclear envelopes or NPCs. Our data suggest that CAN is critical to cell cycle progression and required for both nuclear protein import and mRNA export.
Department of Genetics, St Jude Children's Research Hospital, Memphis, TN 38105, USA.
J
J
van Deursen
J
J
Boer
L
L
Kasper
G
G
Grosveld
D000818:Animals; D001692:Biological Transport; D001755:Blastocyst / Q000166:cytology*; D002467:Cell Nucleus / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect; D016195:G2 Phase; D051379:Mice; D008817:Mice, Mutant Strains; D028861:Nuclear Pore Complex Proteins; D009687:Nuclear Proteins / Q000502:physiology*; D015513:Oncogene Proteins / Q000502:physiology*; D000090063:Proto-Oncogene Mas; D012333:RNA, Messenger / Q000378:metabolism
C000715470:MAS1 protein, human; D028861:Nuclear Pore Complex Proteins; D009687:Nuclear Proteins; C490093:Nup214 protein, mouse; D015513:Oncogene Proteins; D000090063:Proto-Oncogene Mas; D012333:RNA, Messenger