Differential regulation of NF-κB-dependent genes by phosphorylation of Ser-316, -529, and -536.
A–C, microarray data show the different gene regulation effects by different p65 serine to alanine mutants, including S316A, S529A and S536A single mutants and S316/529, S316/536, S529/536 double mutants as well as the S316A/S529A/S536A triple mutant. In this experiment, the data showing the ratio of gene expression levels from mutant(s) p65 versus wtp65, the levels of gene expression from wtp65 sample were considered as 1. Overall, each single mutant was able to decrease 70∼80% that of NF-κB target gene expression. Double and triple mutants had slightly more dramatic effect than each single site mutant. D and E, phosphorylation of Ser-316, Ser-529, and Ser-536 can either commonly (all three sites) regulate some of NF-κB target genes or solely regulate their specific pools of genes. Moreover, two different sites, i.e. Ser-316/529, or Ser-316/536 or Ser-529/536 can commonly regulate some genes, which are not regulated by the third S site, suggesting a hieratical regulation pattern.