title: nup1 mutants exhibit pleiotropic defects in nuclear pore complex function.
abstract: The NUP1 gene of Saccharomyces cerevisiae encodes one member of a family of nuclear pore complex proteins (nucleoporins) conserved from yeast to vertebrates. We have used mutational analysis to investigate the function of Nup1p. Deletion of either the amino- or carboxy-terminal domain confers a lethal phenotype, but partial truncations at either end affect growth to varying extents. Amino-terminal truncation causes mislocalization and degradation of the mutant protein, suggesting that this domain is required for targeting Nup1p to the nuclear pore complex. Carboxy-terminal mutants are stable but do not have wild-type function, and confer a temperature sensitive phenotype. Both import of nuclear proteins and export of poly(A) RNA are defective at the nonpermissive temperature. In addition, nup1 mutant cells become multinucleate at all temperatures, a phenotype suggestive of a defect in nuclear migration. Tubulin staining revealed that the mitotic spindle appears to be oriented randomly with respect to the bud, in spite of the presence of apparently normal cytoplasmic microtubules connecting one spindle pole body to the bud tip. EM analysis showed that the nuclear envelope forms long projections extending into the cytoplasm, which appear to have detached from the bulk of the nucleus. Our results suggest that Nup1p may be required to retain the structural integrity between the nuclear envelope and an underlying nuclear scaffold, and that this connection is required to allow reorientation of the nucleus in response to cytoskeletal forces.
authors.affiliation: Howard Hughes Medical Institute, Department of Genetics, Duke University Medical Center, Durham, North Carolina 27710.
authors.forename: A M
authors.initials: AM
authors.lastname: Bogerd
authors.forename: J A
authors.initials: JA
authors.lastname: Hoffman
authors.forename: D C
authors.initials: DC
authors.lastname: Amberg
authors.forename: G R
authors.initials: GR
authors.lastname: Fink
authors.forename: L I
authors.initials: LI
authors.lastname: Davis
mesh_terms: D001483:Base Sequence; D001692:Biological Transport; D002467:Cell Nucleus / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure; D004252:DNA Mutational Analysis; D005656:Fungal Proteins / Q000378:metabolism; D005800:Genes, Fungal; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009685:Nuclear Envelope / Q000502:physiology* / Q000648:ultrastructure; D028861:Nuclear Pore Complex Proteins; D009687:Nuclear Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000502:physiology*; D010641:Phenotype; D012331:RNA, Fungal / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000502:physiology* / Q000648:ultrastructure; D013696:Temperature; D014404:Tubulin / Q000032:analysis
chemical_list: D005656:Fungal Proteins; D008565:Membrane Proteins; C064550:NUP1 protein, S cerevisiae; D028861:Nuclear Pore Complex Proteins; D009687:Nuclear Proteins; D012331:RNA, Fungal; D012333:RNA, Messenger; D014404:Tubulin