Regulation of Stat3 nuclear import by importin alpha5 and importin alpha7 via two different functional sequence elements.
Regulated import of STAT proteins into the nucleus through the nuclear pores is a vital event. We previously identified Arg214/215 in the coiled-coil domain and Arg414/417 in the DNA binding domain involved in the ligand-induced nuclear translocation of Stat3. In this study, we investigated the mechanism for Stat3 nuclear transport. We report here that among five ubiquitously expressed human importin alphas, importin alpha5 and alpha7, but not importin alpha1, alpha3, and alpha4, bind to Stat3 upon cytokine stimulation. Similar results were observed for Stat1, but not for Stat5a and 5b, which were unable to interact with any of the importin alphas. The C-terminus of importin alpha5 is necessary but not sufficient for Stat3 binding. Truncation mutant of Stat3 (aa1-320) that contains Arg214/215 exhibits specific binding to importin alpha5, and an exclusive nuclear localization. Point mutations of Arg214/215 in this mutant destroy importin alpha5 binding and its nuclear localization. In contrast, the truncation mutant (aa320-770) including Arg414/417 fails to interact with importin alpha5 and is localized in the cytoplasm. However, both sequence elements are necessary for the full-length Stat3's interaction with importin alpha5. These results suggest that Arg214/215 is likely the binding site for importin alpha5, whereas Arg414/417 may not be involved in the direct binding, but necessary for maintaining the proper conformation of Stat3 dimer for importin binding. A model for Stat3 nuclear translocation is proposed based on these data.
Ma
Jing
J
Signal Transduction Laboratory, Institute of Molecular and Cell Biology, Proteos Building, 61 Biopolis Drive, Singapore, 138673, Republic of Singapore.
Cao
Xinmin
X
D021581:Active Transport, Cell Nucleus; D000595:Amino Acid Sequence; D000818:Animals; D001120:Arginine / Q000378:metabolism; D019556:COS Cells; D002467:Cell Nucleus / Q000378:metabolism*; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D006801:Humans; D051379:Mice; D008954:Models, Biological; D009154:Mutation / Q000235:genetics; D019000:Phosphotyrosine / Q000378:metabolism; D011485:Protein Binding; D025941:Protein Interaction Mapping; D050794:STAT1 Transcription Factor / Q000378:metabolism; D050796:STAT3 Transcription Factor / Q000737:chemistry / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D028901:alpha Karyopherins / Q000737:chemistry / Q000378:metabolism*
C423142:KPNA1 protein, human; C494630:KPNA6 protein, human; D050794:STAT1 Transcription Factor; D050796:STAT3 Transcription Factor; D028901:alpha Karyopherins; D019000:Phosphotyrosine; D001120:Arginine