NIP1, a gene required for nuclear transport in yeast

Proc Natl Acad Sci U S A. 1992 Nov 1;89(21):10355-9. doi: 10.1073/pnas.89.21.10355.

Abstract

Cytochrome c with a nuclear localization signal added at the N terminus was mistargeted to the nucleus, resulting in a yeast strain deficient in mitochondrial cytochrome c. Reversion of this strain allowed the isolation of temperature-conditional mutants defective in nuclear transport, as demonstrated with one of these mutants, nip1-1, that was shown to be defective in nuclear accumulation of a LacZ protein containing a nuclear localization signal of the yeast ribosomal protein L29. The NIP1+ gene was cloned and shown to encode a 93,143-Da protein. Furthermore, an epitope-labeled NIP1 protein migrated in SDS/polyacrylamide gels with a mass of approximately 100,000 Da and was shown by immunofluorescence to localize mainly in the cytoplasm. NIP1+ was shown to be an essential gene by gene disruption experiments. Intriguingly, NIP1 has a serine-rich acidic N-terminal region that is similar in this regard to the N-terminal region of a previously described nuclear localization signal-binding protein, NSR1.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Alleles
  • Amino Acid Sequence
  • Base Sequence
  • Cell Nucleus / metabolism*
  • Cloning, Molecular
  • Codon / genetics
  • Cytochrome c Group / genetics*
  • Eukaryotic Initiation Factor-3
  • Fungal Proteins / genetics*
  • Fungal Proteins / isolation & purification
  • Genes, Fungal*
  • Genetic Complementation Test
  • Immunoblotting
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Nuclear Proteins / genetics*
  • Nuclear Proteins / isolation & purification
  • Phenotype
  • Protein Biosynthesis
  • Recombinant Fusion Proteins / isolation & purification
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae Proteins*
  • TATA Box
  • beta-Galactosidase / genetics
  • beta-Galactosidase / isolation & purification

Substances

  • Codon
  • Cytochrome c Group
  • Eukaryotic Initiation Factor-3
  • Fungal Proteins
  • NIP1 protein, S cerevisiae
  • Nuclear Proteins
  • Recombinant Fusion Proteins
  • Saccharomyces cerevisiae Proteins
  • beta-Galactosidase

Associated data

  • GENBANK/L02899