The significance of phosphorylation for FXR. Chlorocebus sabaeus

The serine or threonine between two zinc fingers of the DNA binding domains (DBD) is highly conserved in a majority of nuclear receptors. In the present study, we focused on the serine 154 of farnesoid X receptor (FXR). To determine the influence of phosphomimetic or non-phosphomimetic mutation for FXR on gene expressions, we employed cDNA microarray analysis using COS-1 cells which ectopically express FXR S154A or FXR S154D with DMSO or CDCA. Overall design: COS-1 cells were transfected with human FXR S154A mutant or its S154D for 24 hr. The cells were subsequently incubated with 0.1%DMSO or 50 mMCDCA for 24 hr. Three replicates were used for each treatment group.

DNA结合结构域（DNA binding domain, DBD）的两个锌指之间的丝氨酸或苏氨酸残基，在大多数核受体中均高度保守。本研究聚焦于法尼醇X受体（farnesoid X receptor, FXR）的第154位丝氨酸残基。为探明FXR的磷酸模拟突变与非磷酸模拟突变对基因表达的影响，我们以分别经二甲基亚砜（DMSO）或鹅脱氧胆酸（CDCA）处理的、异位表达FXR S154A或FXR S154D的COS-1细胞为材料，开展cDNA微阵列分析。
整体实验设计：将转染了人源FXR S154A突变体或S154D突变体的COS-1细胞培养24小时，随后分别以0.1% DMSO或50 mM CDCA处理24小时，每个处理组均设置3次生物学重复。