Dissection of amyloid-beta precursor protein-dependent transcriptional transactivation.
Amyloid-beta precursor protein (APP) forms a transcriptionally active complex with the adaptor protein Fe65 and the histone acetyltransferase Tip60, but the mechanism of transcriptional activation that is mediated by APP and Fe65 remains unclear. APP is cleaved by gamma-secretase similar to Notch, whose intracellular domain activates transcription by interacting with nuclear transcription factors. To test whether the APP intracellular domain (AICD) functions analogously, we investigated how APP and Fe65 transactivate a Gal4 fusion protein of Tip60. Consistent with the Notch paradigm, we observe that gamma-cleavage of APP and nuclear translocation of Fe65 are required for transactivation. Surprisingly, however, we find that nuclear translocation of the AICD may be dispensable and that only membrane-tethered AICD (i.e. AICD coupled to a transmembrane region) and not free AICD (i.e. soluble AICD) is a potent transactivator of transcription. Membrane-tethered AICD recruits Fe65 and mediates the activation of bound Fe65 that is then released for nuclear translocation by gamma-cleavage together with the AICD. Our data suggest that transcriptional transactivation by APP and Notch may involve distinct mechanisms; whereas the Notch intracellular domain directly functions in the nucleus, the AICD acts indirectly by activating Fe65.
Cao
Xinwei
X
Center for Basic Neuroscience, Department of Molecular Genetics, and Howard Hughes Medical Institute, The University of Texas Southwestern Medical Center, Dallas, Texas 75390-9111, USA.
Südhof
Thomas C
TC
D000123:Acetyltransferases / Q000378:metabolism; D021581:Active Transport, Cell Nucleus; D016564:Amyloid beta-Protein Precursor / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D001665:Binding Sites; D019556:COS Cells; D002462:Cell Membrane / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation; D006367:HeLa Cells; D051548:Histone Acetyltransferases; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D000074443:Lysine Acetyltransferase 5; D008565:Membrane Proteins / Q000378:metabolism; D008954:Models, Biological; D009154:Mutation; D009419:Nerve Tissue Proteins / Q000378:metabolism; D009687:Nuclear Proteins / Q000378:metabolism; D010957:Plasmids / Q000378:metabolism; D011233:Precipitin Tests; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D051381:Rats; D051880:Receptors, Notch; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D014158:Transcription, Genetic; D015533:Transcriptional Activation
C091042:APBB1 protein, human; D016564:Amyloid beta-Protein Precursor; C486242:Apbb1 protein, rat; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D009687:Nuclear Proteins; D051880:Receptors, Notch; D011993:Recombinant Fusion Proteins; D000123:Acetyltransferases; D051548:Histone Acetyltransferases; C098489:KAT5 protein, human; D000074443:Lysine Acetyltransferase 5