Intracellular distribution of a nuclear localization signal binding protein.
The transport of proteins into the nucleus requires the recognition of a nuclear localization signal sequence. Several proteins that interact with these sequences have been identified, including one of about 66 kDa. We have prepared antibodies that recognize the 66-kDa nuclear localization signal binding protein (NLSBP) and inhibit nuclear localization in vitro. By immunofluorescence, it is seen that the NLSBP is predominantly cytoplasmic and is distributed peripherally around the nucleus and the microtubule organizing center. There is also a weak punctate staining of the surface of the nucleus. Methanol-fixed cells can also be stained directly with fluorescently labeled karyophilic proteins. These stains reveal the same cytoplasmic structures as anti-NLSBP. The expression of the NLSBP is growth dependent. When cells grown to confluence are examined, the cytoplasmic staining is greatly reduced, leaving the punctate nuclear staining as the predominant feature. In serum-starved cells, very little staining of either the cytoplasm or the nucleus can be seen. Upon simulation by the addition of serum, the original cytoplasmic and nuclear envelope staining is restored. Cells grown in the presence of colchicine or taxol have an altered NLSBP distribution but apparently normal cytoplasmic nuclear transport.
Li
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Department of Biochemistry, New York University School of Medicine, New York 10016.
Shi
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Thomas
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D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology; D001692:Biological Transport; D015153:Blotting, Western; D002455:Cell Division; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism*; D002522:Chlorocebus aethiops; D003078:Colchicine / Q000494:pharmacology; D003593:Cytoplasm / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D006367:HeLa Cells; D006801:Humans; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000276:immunology / Q000378:metabolism*; D017239:Paclitaxel / Q000494:pharmacology; D010750:Phosphoproteins / Q000276:immunology / Q000378:metabolism*; D014982:Xenopus laevis
D000906:Antibodies; C075564:NOLC1 protein, human; D009687:Nuclear Proteins; D010750:Phosphoproteins; D017239:Paclitaxel; D003078:Colchicine