Characterization of a Drosophila phosphorylation-dependent nuclear-localization-signal-binding protein.
A 94 kDa nuclear-localization-signal (NLS)-binding protein was purified from Drosophila embryos. The NLS of the simian-virus-40 T-antigen is specifically bound by the dephosphorylated form of the protein. After phosphorylation, the affinity of the protein for the NLS is sharply decreased. In the dephosphorylated form, p94 (protein of 94 kDa) is the major NLS-binding protein in Drosophila embryos. Immunoprecipitation confirmed the ATP-dependent phosphorylation of p94, and co-precipitation of two additional phosphorylated proteins, indicated that the NLS-binding protein is part of a larger complex in Drosophila embryos. In agreement with the immunoprecipitation results, cross-linking experiments demonstrated the interaction of p94 with three additional proteins. These protein-protein interactions were also phosphorylation-dependent.
Cserpán
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I
Biological Research Center of the Hungarian Academy of Sciences, 6701 Szeged, Hungary.
Máthé
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E
Patthy
A
A
Udvardy
A
A
D000255:Adenosine Triphosphate / Q000378:metabolism / Q000494:pharmacology; D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000378:metabolism; D001667:Binding, Competitive; D002455:Cell Division / Q000502:physiology; D003432:Cross-Linking Reagents / Q000378:metabolism; D004072:Digitonin / Q000494:pharmacology; D004331:Drosophila melanogaster / Q000737:chemistry* / Q000196:embryology; D004591:Electrophoresis, Polyacrylamide Gel; D005029:Ethylenediamines / Q000378:metabolism; D015151:Immunoblotting; D019476:Insect Proteins / Q000302:isolation & purification / Q000378:metabolism; D019913:Nuclear Localization Signals; D009687:Nuclear Proteins / Q000302:isolation & purification / Q000378:metabolism*; D010750:Phosphoproteins / Q000302:isolation & purification / Q000378:metabolism*; D010766:Phosphorylation; D011233:Precipitin Tests; D011485:Protein Binding; D013388:Succinimides / Q000378:metabolism
D000952:Antigens, Polyomavirus Transforming; D003432:Cross-Linking Reagents; D005029:Ethylenediamines; D019476:Insect Proteins; C075564:NOLC1 protein, human; D019913:Nuclear Localization Signals; D009687:Nuclear Proteins; D010750:Phosphoproteins; D013388:Succinimides; C051801:ethyleneglycol bis(succinimidyl succinate)hydroxyethylethylenediamine triacetic acid; D000255:Adenosine Triphosphate; D004072:Digitonin