Identification of four nuclear transport signal-binding proteins that interact with diverse transport signals.
The transport of proteins into the nucleus requires not only the presence of a nuclear transport signal on the targeted protein but also the signal recognition proteins and the nuclear pore translocation apparatus. Complicating the search for the signal recognition proteins is the fact that the nuclear transport signals identified share little obvious homology. In this study, synthetic peptides homologous to the nuclear transport signals from the simian virus 40 large T antigen, Xenopus oocyte nucleoplasmin, adenovirus E1A, and Saccharomyces cerevisiae MAT alpha 2 proteins were coupled to a UV-photoactivable cross-linker and iodinated for use in an in vitro cross-linking reaction with cellular lysates. Four proteins, p140, p100, p70, and p55, which specifically interacted with the nuclear transport signal peptides were identified. Unique patterns of reactivity were observed with closely related pairs of nuclear transport signal peptides. Competition experiments with labeled and unlabeled peptides demonstrated that heterologous signals were able to bind the same protein and suggested that diverse signals use a common transport pathway. The subcellular distribution of the four nuclear transport signal-binding proteins suggested that nuclear transport involves both cytoplasmic and nuclear receptors. The four proteins were not bound by wheat germ agglutinin and were not associated tightly with the nuclear pore complex.
Yamasaki
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L
Department of Virology and Immunology, Southwest Foundation for Biomedical Research, San Antonio, Texas.
Kanda
P
P
Lanford
R E
RE
D017456:Adenovirus Early Proteins; D000595:Amino Acid Sequence; D000818:Animals; D000957:Antigens, Viral, Tumor / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism*; D003432:Cross-Linking Reagents; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009687:Nuclear Proteins / Q000378:metabolism; D056489:Nucleoplasmins; D009856:Oncogene Proteins, Viral / Q000378:metabolism; D010750:Phosphoproteins; D011485:Protein Binding; D021382:Protein Sorting Signals / Q000138:chemical synthesis / Q000378:metabolism*; D051381:Rats; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D015398:Signal Transduction; D013539:Simian virus 40 / Q000378:metabolism; D014466:Ultraviolet Rays; D014909:Wheat Germ Agglutinins
D017456:Adenovirus Early Proteins; D000957:Antigens, Viral, Tumor; D003432:Cross-Linking Reagents; D009687:Nuclear Proteins; D056489:Nucleoplasmins; D009856:Oncogene Proteins, Viral; D010750:Phosphoproteins; D021382:Protein Sorting Signals; D014909:Wheat Germ Agglutinins