PPARγ and NF-κB regulate the gene promoter activity of their shared repressor, TNIP1

Human TNFAIP3 interacting protein 1 (TNIP1) has diverse functions including support of HIV replication through its interaction with viral Nef and matrix proteins, reduction of TNFα-induced signaling through its interaction with NF-κB pathway proteins, and corepression of agonist-bound retinoic acid receptors and peroxisome proliferator-activated receptors (PPAR). The wide tissue distribution of TNIP1 provides the opportunity to influence numerous cellular responses in these roles and defining control of TNIP1 expression would be central to improved understanding of its impact on cell function. We cloned 6 kb of the human TNIP1 promoter and performed predictive and functional analyses to identify regulatory elements. The promoter region proximal to the transcription start site is GC-rich without a recognizable TATA box. In contrast to this proximal ~ 500 bp region, 6 kb of the promoter increased reporter construct constitutive activity over five-fold. Throughout the 6 kb length, in silico analysis identified several potential binding sites for both constitutive and inducible transcription factors; among the latter were candidate NF-κB binding sequences and peroxisome proliferator response elements (PPREs). We tested NF-κB and PPAR regulation of the endogenous TNIP1 gene and cloned promoter by expression studies, electrophoretic mobility shift assays, and chromatin immunoprecipitations. We validated NF-κB sites in the TNIP1 promoter proximal and distal regions as well as one PPRE in the distal region. The ultimate control of the TNIP1 promoter is likely to be a combination of constitutive transcription factors and those subject to activation such as NF-κB and PPAR.

► Six kilobases of the human TNIP1 promoter were cloned and examined in silico. ► Functional NF-κB and PPAR sites have been identified in the human TNIP1 promoter. ► NF-κB and PPARγ bind to the endogenous TNIP1 promoter. ► The TNIP1 promoter PPRE functions in an orientation-independent manner. ► TNIP1 expression is likely to vary as abundance and activity of NF-κB and PPAR change.