Characterization of mouse retinoid X receptor (RXR)-beta gene promoter: negative regulation by tumor necrosis factor...

A genomic clone of mouse retinoid X receptor (RXR)-beta (Rxrb) has recently been isolated and mapped within the H2-K region of the mouse major histocompatibility complex. A putative 250-bp promoter, which is located between Rxrb and H2-Ke4, and may possibly be their common promoter, has also been identified. In order to study the gene regulation of Rxrb, we analyzed the transcriptional function of the Rxrb promoter with chimeric constructs containing the Rxrb promoter fragments fused upstream of a firefly luciferase cDNA, which were transiently transfected into rat GH3 cells. We found that 1) a part of the H2-Ke4 genomic region (1.9-kb), as well as the 250-bp promoter, was transcriptionally active as an Rxrb promoter; 2) tumor necrosis factor (TNF)-alpha significantly repressed the activity of the 250-bp promoter although thyroid hormone, 9-cis retinoic acid, interleukin (IL)-1beta, and IL-6 did not affect the activity; 3) either the change in orientation or point mutations of a consensus NF-kappaB site located in the 250-bp promoter did not affect the repression; 4) SB 203580, a highly specific inhibitor of p38 mitogen-activated protein (MAP) kinase, completely abolished the repression by TNF-alpha. These data suggest that TNF-alpha represses the promoter activity of the 250-bp region, and the repression is mediated by p38 MAP kinase independent of NF-kappaB. We thus have first shown a relation between the retinoic acid receptor and a cytokine TNF-alpha.