A role of uncoupling protein 2 (UCP2) as negative modulator of insulin secretion has been suggested but the transcriptional pathways regulating -cell UCP2 gene expression have been established in rodents only. We show here that the underlying sequence motifs are not conserved in the human gene and provide evidence for regulatory mechanisms involving the transcriptional cofactor peroxisome proliferator-activated receptor coactivator-1 (PGC-1). PGC-1 potentiates thyroid hormone (T3) mediated transcriptional activation of the human UCP2 gene in INS-1E cells. Two thyroid hormone response elements (TREs) located at -322/-317 (TRE1) and -170/-165 (TRE2) were identified and mutation of either TRE1 or TRE2 abrogated the stimulatory effect of T3 treatment. Furthermore two E-Box motifs at -911/-906 (E1) and -743/-738 (E2) are involved in the regulation of UCP2 gene expression by sterol regulatory element binding protein isoforms SREBP-1a, -1c and -2. Mutational analysis revealed that the presence of either E1 or E2 is sufficient to mediate activation of UCP2 gene transcription by nuclear active SREBPs. PGC-1 coactivates LXR mediated expression of SREBP-1c as well as dexamethasone stimulated SREBP-2 expression in INS-1E cells. These transcriptional responses are antagonized by orphan nuclear receptor short heterodimer partner (SHP) overexpression which might explain its positive effects on GSIS in -cells overexpressing UCP2. We also provide evidence that despite a lack of sequence homology within the regulatory region the principle mechanisms regulating UCP2 gene expression are similar in rats and humans being consistent with a role for UCP2 as a modulator of insulin secretion in humans.