期刊名称：Proceedings of the National Academy of Sciences
印刷版ISSN：0027-8424
电子版ISSN：1091-6490
出版年度：2013
卷号：110
期号：5
页码：E377-E386
DOI：10.1073/pnas.1212990110
语种：English
出版社：The National Academy of Sciences of the United States of America
摘要：Chemically induced dimerization is an important tool in chemical biology for the analysis of protein function in cells. Here we report the use of the natural product fusicoccin (FC) to induce dimerization of 14-3-3-fused target proteins with proteins tagged to the C terminus (CT) of the H+-ATPase PMA2. To prevent nonproductive or detrimental interactions of the 14-3-3 proteins and CT fusions with endogenous cell proteins, their interaction surface was engineered to facilitate FC-induced dimerization exclusively between the introduced protein constructs. Live-cell imaging documented the reversible FC-induced translocation of 14-3-3 and CT to different cell compartments depending on localization sequences fused to their dimerization partner protein. The functionality of this system was demonstrated by the FC-induced importation of the NF-{kappa}B-CT into the nucleus. In HeLa cells, FC-mediated dimerization of the NF-{kappa}B-CT with a constitutively nuclear-localized 14-3-3 protein led to an NF-{kappa}B-specific cellular response by inducing IL-8 secretion.
