title: Specific binding of nuclear localization sequences to plant nuclei.
abstract: We have begun to dissect the import apparatus of higher plants by examining the specific association of nuclear localization sequences (NLSs) with purified plant nuclei. Peptides to the simian virus 40 (SV40) large T antigen NLS and a bipartite NLS of maize were allowed to associate with tobacco and maize nuclei. Wild-type NLSs were found to compete for a single class of low-affinity binding sites having a dissociation constant (Kd) of approximately 200 microM. Peptides to mutant NLSs, which are inefficient in stimulating import, were poor competitors, as were reverse wild-type and non-NLS peptides. The NLS binding site was proteinaceous and resistant to extraction under conditions where pores were still associated. In addition, immunofluorescence and immunoelectron microscopy indicated that binding was at the nuclear envelope. Overall, plant nuclei may be an excellent system to identify components of the import apparatus.
authors.affiliation: Department of Energy Plant Research Laboratory, Michigan State University, East Lansing 48824-1312.
authors.forename: G R
authors.initials: GR
authors.lastname: Hicks
authors.forename: N V
authors.initials: NV
authors.lastname: Raikhel
mesh_terms: D000595:Amino Acid Sequence; D000957:Antigens, Viral, Tumor / Q000235:genetics / Q000378:metabolism; D001665:Binding Sites; D001667:Binding, Competitive; D002467:Cell Nucleus / Q000378:metabolism; D008969:Molecular Sequence Data; D019913:Nuclear Localization Signals; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D010944:Plants / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D010947:Plants, Toxic; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism; D003313:Zea mays / Q000235:genetics / Q000378:metabolism
chemical_list: D000957:Antigens, Viral, Tumor; D019913:Nuclear Localization Signals; D009687:Nuclear Proteins; D010940:Plant Proteins