Nicotinamide adenine dinucleotide (NAD) plays a critical role in the maintenance of cellular energy homeostasis. alpha-Amino-beta-carboxymuconate-epsilon-semialdehyde decarboxylase (ACMSD) is the key enzyme regulating de novo synthesis of NAD from l-tryptophan (Trp), designated the Trp-NAD pathway. Acmsd gene expression was found to be under the control of both hepatocyte nuclear factor 4alpha (HNF4alpha) and peroxisome proliferator-activated receptor alpha (PPARalpha). Constitutive expression of ACMSD mRNA levels were governed by HNF4alpha and downregulated by activation of PPARalpha by the ligand Wy-14,643 ([4-chloro-6-(2,3-xylidino)-2-pyrimidinylthio]acetic acid]), as revealed by studies with hepatic HNF4alpha-null mice and PPARalpha-null mice, respectively. Transient transfection and electrophoretic mobility shift analyses showed an HNF4alpha binding site in the Acmsd gene promoter that directed transactivation of reporter gene constructs by HNF4alpha. The Acmsd promoter was not responsive to PPARalpha in transactivation assays. Wy-14,643 treatment decreased HNF4alpha protein levels in wild-type, but not PPARalpha-null, mouse livers, with no changes in HNF4alpha mRNA. These results show that Wy-14,643, through PPARalpha, post-transcriptionally down-regulates HNF4alpha protein levels, leading to reduced expression of the HNF4alpha target gene Acmsd.