Interaction of Estrogen Receptor-α Transactivation Domain With Nuclear Proteins of Mouse Brain: p68 RNA Helicase Shows Age- and Sex-Specific Change

Estrogen receptor (ER)-alpha interacts with nuclear proteins to mediate its multiple functions in the brain. However, it is not known which proteins interact with the ER alpha-transactivation domain (TAD) in mouse brain and whether they change with age and sex. Therefore, we have used affinity-purified GST-tagged mouse ER alpha-TAD fusion protein for interaction with nuclear proteins from the mouse brain. The pull-down assay and far-Western blotting detected four nuclear proteins of 100, 80, 68, and 50 kD. We have recently identified the 80-kD protein as MTA1 and demonstrated its decrease in old age. Here we report alteration in the interaction and expression of the 68-kD protein of adult and old mice of both sexes. This protein was identified as p68 RNA helicase through NCBI database search, immunoprecipitation, and immunoblotting. Further analysis showed that the extent of its interaction was relatively lower in old mice of both sexes and in male mice of both ages compared with their counterparts. However, the expression of p68 was significantly lower in old males compared with adult males, although other groups did not show significant changes. Such age- and sex-specific interaction of p68 suggests its implication in ER alpha-mediated brain functions during aging. (C) 2008 Wiley-Liss, Inc.