A general strategy for inactivation of target proteins is presented, which we have termed "oligomerization chain reaction." This technique is based on the fusion of the self-associating coiled-coil (CC) domain of the nuclear factor promyelocytic leukemia (PML) to target proteins that are able to self-associate naturally. Oligomerization through the CC region of promyelocytic leukemia, and through the natural self-associating domain, triggers the oligomerization chain reaction, leading to formation of large molecular weight complexes and functional inactivation of the target. As a test case, we have chosen the oncosuppressor p53, naturally occurring as a tetramer. Fusion of the CC to p53 leads to formation of stable high molecular weight complexes - as shown by size exclusion chromatography - to which wild-type p53 is recruited with high efficiency. CC-p53 chimeras delocalize wild-type p53 to the cytoplasm and inhibit its transcriptional regulatory properties, resulting in a loss of p53 function. We propose that this strategy may be of general application to self-associating factors and represent a complementary approach to currently used functional inactivation-based strategies.
|Titolo:||Targeting protein inactivation through an oligomerization chain reaction|
PELICCI, PIER GIUSEPPE (Penultimo)
|Parole Chiave:||3T3 Cells; Animals; Tumor Suppressor Protein p53; Humans; Transcription, Genetic; Recombinant Fusion Proteins; Mice; Cell Nucleus; Plasmids; Models, Biological; Protein Binding; Tumor Suppressor Proteins; Microscopy, Fluorescence; DNA, Complementary; Neoplasm Proteins; Tumor Cells, Cultured; Transcription Factors; Nuclear Proteins; Transfection; Cytoplasm; Protein Structure, Tertiary|
|Settore Scientifico Disciplinare:||Settore MED/04 - Patologia Generale|
|Data di pubblicazione:||19-feb-2002|
|Digital Object Identifier (DOI):||10.1073/pnas.042460299|
|Appare nelle tipologie:||01 - Articolo su periodico|