ArticleThe Journal of cell biology1993
Reconstitution of nuclear protein transport with semi-intact yeast cells.
Article in The Journal of cell biology, 1993. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 39 papers.
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Who cites it
39 citing papers in PubMed, 86 citations in OpenAlex.
- An integrated machine-learning model to predict nucleosome architecture.Nucleic acids research · 2024Article
- The Orf9b protein of SARS-CoV-2 modulates mitochondrial protein biogenesis.The Journal of cell biology · 2023Article
- Impact of DNA methylation on 3D genome structure.Nature communications · 2021Article
- Calmodulin-driven nuclear entry: trigger for sex determination and terminal differentiation.The Journal of biological chemistry · 2009Review
- Replication of Carnation Italian ringspot virus defective interfering RNA in Saccharomyces cerevisiae.Journal of virology · 2003Article
- The Nsp1p carboxy-terminal domain is organized into functionally distinct coiled-coil regions required for assembly of nucleoporin subcomplexes and nucleocytoplasmic transport.Molecular and cellular biology · 2001Article
- Mechanisms controlling subcellular localization of the G(1) cyclins Cln2p and Cln3p in budding yeast.Molecular and cellular biology · 2001Article
- An exclusively nuclear RNA-binding protein affects asymmetric localization of ASH1 mRNA and Ash1p in yeast.The Journal of cell biology · 2001Article
- The heat shock protein Ssa2p is required for import of fructose-1, 6-bisphosphatase into Vid vesicles.The Journal of cell biology · 2000Article
- Brome mosaic virus RNA replication proteins 1a and 2a colocalize and 1a independently localizes on the yeast endoplasmic reticulum.Journal of virology · 1999Article
- Interactions between a nuclear transporter and a subset of nuclear pore complex proteins depend on Ran GTPase.Molecular and cellular biology · 1999Article
- The nuclear pore complex.Plant molecular biology · 1998Review
- Yeast heat shock mRNAs are exported through a distinct pathway defined by Rip1p.Genes & development · 1997Article
- Hrp1, a sequence-specific RNA-binding protein that shuttles between the nucleus and the cytoplasm, is required for mRNA 3'-end formation in yeast.Genes & development · 1997Article
- Nuclear localization signal binding protein from Arabidopsis mediates nuclear import of Agrobacterium VirD2 protein.Proceedings of the National Academy of Sciences of the United States of America · 1997Article
- Nucleocytoplasmic transport of macromolecules.Microbiology and molecular biology reviews : MMBR · 1997Review
- The vertebrate GLFG nucleoporin, Nup98, is an essential component of multiple RNA export pathways.The Journal of cell biology · 1997Article
- In vitro reconstitution of a heterotrimeric nucleoporin complex consisting of recombinant Nsp1p, Nup49p, and Nup57p.Molecular biology of the cell · 1997Article
- GLE2, a Saccharomyces cerevisiae homologue of the Schizosaccharomyces pombe export factor RAE1, is required for nuclear pore complex structure and function.Molecular biology of the cell · 1996Article
- Nuclear transport defects and nuclear envelope alterations are associated with mutation of the Saccharomyces cerevisiae NPL4 gene.Molecular biology of the cell · 1996Article
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
We have developed an in vitro nuclear protein import reaction from semi-intact yeast cells. The reaction uses cells that have been permeabilized by freeze-thaw after spheroplast formation. Electron microscopic analysis and antibody-binding experiments show that the nuclear envelope remains intact but the plasma membrane is perforated. In the presence of ATP and cytosol derived from yeast or mammalian cells, a protein containing the nuclear localization sequence (NLS) of SV40 large T-antigen is transported into the nucleus. Proteins with mutant NLSs are not imported. In the absence of cytosol, binding of NLS-containing proteins occurs at the nuclear envelope. N-ethylmaleimide treatment of the cytosol as well as antibodies to the nuclear pore protein Nsp1 inhibit import but not binding to the nuclear envelope. Yeast mutants defective in nuclear protein transport were tested in the in vitro import reaction. Semi-intact cells from temperature-sensitive nsp1 mutants failed to import but some binding to the nuclear envelope was observed. On the other hand, no binding and thus no import into nuclei was observed in semi-intact nsp49 cells which are mutated in another nuclear pore protein. Np13 mutants, which are defective for nuclear protein import in vivo, were also deficient in the binding step under the in vitro conditions. Thus, the transport defect in these mutants is at the level of the nucleus and the point at which nuclear transport is blocked can be defined.
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