The mechanism of ran import into the nucleus by nuclear transport factor 2.
The small GTPase Ran is essential for virtually all nucleocytoplasmic transport events. It is hypothesized that Ran drives vectorial transport of macromolecules into and out of the nucleus via the establishment of a Ran gradient between the cytoplasm and nucleoplasm. Although Ran shuttles between the nucleus and cytoplasm, it is concentrated in the nucleus at steady state. We show that nuclear transport factor 2 (NTF2) is required to concentrate Ran in the nucleus in the budding yeast, Saccharomyces cerevisiae. To analyze the mechanism of Ran import into the nucleus by NTF2, we use mutants in a variety of nuclear transport factors along with biochemical analyses of NTF2 complexes. We find that Ran remains concentrated in the nucleus when importin-mediated protein import is disrupted and demonstrate that NTF2 does not form a stable complex with the transport receptor, importin-beta. Consistent with a critical role for NTF2 in establishing and maintaining the Ran gradient, we show that NTF2 is required for early embryogenesis in Caenorhabditis elegans. Our data distinguish between two possible mechanisms for Ran import by NTF2 and demonstrate that Ran import is independent from importin-beta-mediated protein import.
Quimby
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BB
Department of Biochemistry, Emory University School of Medicine and the Department of Biology, Emory University, Atlanta, Georgia 30322, USA.
Lamitina
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T
L'Hernault
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Corbett
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AH
D000595:Amino Acid Sequence; D001692:Biological Transport; D002352:Carrier Proteins / Q000737:chemistry / Q000502:physiology*; D002467:Cell Nucleus / Q000378:metabolism*; D028884:Karyopherins; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000737:chemistry / Q000502:physiology*; D029543:Nucleocytoplasmic Transport Proteins; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D020931:ran GTP-Binding Protein / Q000378:metabolism*
D002352:Carrier Proteins; D028884:Karyopherins; D009687:Nuclear Proteins; D029543:Nucleocytoplasmic Transport Proteins; D020931:ran GTP-Binding Protein