Abstract
Chromosome maintenance region 1 (CRM1), a protein that shares sequence similarities with the karyopherin beta family of proteins involved in nuclear import pathway, was shown to form a complex with the leucine-rich nuclear export signal (NES). This interaction was inhibited by leptomycin B, a drug that prevents the function of the CRM1 protein in yeast. To analyze the role of the CRM1-NES interaction in nuclear export, a transport assay based on semipermeabilized cells was developed. In this system, which reconstituted NES-, cytosol-, and energy-dependent nuclear export, leptomycin B specifically blocked export of NES-containing proteins. Thus, the CRM1 protein could act as a NES receptor involved in nuclear protein export.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adenosine Triphosphate / metabolism
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Amino Acid Sequence
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Biological Transport / drug effects
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Carrier Proteins / physiology*
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Cell Nucleus / metabolism*
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DNA-Binding Proteins / chemistry
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DNA-Binding Proteins / metabolism*
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Exportin 1 Protein
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Fatty Acids, Unsaturated / pharmacology
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Fluorescent Antibody Technique, Indirect
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HeLa Cells
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Humans
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I-kappa B Proteins*
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Immunoblotting
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Karyopherins*
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NF-KappaB Inhibitor alpha
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Nuclear Localization Signals
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Nuclear Proteins / metabolism*
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Protein Sorting Signals / chemistry
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Protein Sorting Signals / metabolism*
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Pyruvate Kinase / metabolism
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Receptors, Cytoplasmic and Nuclear*
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Recombinant Fusion Proteins / metabolism
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Transfection
Substances
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Adenosine Triphosphate
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Carrier Proteins
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DNA-Binding Proteins
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Fatty Acids, Unsaturated
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I-kappa B Proteins
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Karyopherins
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NF-KappaB Inhibitor alpha
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Nuclear Localization Signals
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Nuclear Proteins
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Protein Sorting Signals
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Pyruvate Kinase
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Receptors, Cytoplasmic and Nuclear
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Recombinant Fusion Proteins
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Exportin 1 Protein
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NFKBIA protein, human
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leptomycin B