Biochemical and Biophysical Research Communications, Vol.348, No.1, 13-24, 2006
Nuclear localization of coactivator RAC3 is mediated by a bipartite NLS and importin alpha 3
The nuclear receptor coactivator RAC3 (also known as SRC-3/ACTR/AIB1/p/CIP/TRAM-1) belongs to the p160 coactivator family, which are involved in several physiological processes and diseases. Here we have investigated how RAC3 is translocated into the nucleus and show that it is mediated through a bipartite NLS and importin alpha 3. This bipartite NLS is located within the conserved bHLH domain, and its mutation abolished nuclear localization. The NLS is also sufficient to cause nuclear import of EGFP, and the activity requires basic amino acids within the NLS. RAC3 binds strongly to importin 0, which also depends on the basic amino acids. Functionally, RAO cytoplasmic mutant loses its ability to enhance transcription, suggesting that nuclear localization is essential for coactivator function. Together, these results reveal a previous unknown mechanism for nuclear translocation of p 160 coactivators and a critical function of the conserved bHLH within the coactivator. (c) 2006 Elsevier Inc. All rights reserved.