Apo2L, or TRAIL, is a type II integral membrane protein belonging to the TNF family which induces apoptotic cell death in a variety of human tumor cells. Apo2L is expressed in many tissues, suggesting that it is nontoxic to normal cells. We found that Apo2L mRNA was induced by interferon (IFN)-alpha and -beta, but not -gamma, in Jurkat cells. To gain a better understanding of the molecular mechanisms that regulate expression of Apo2L, we have characterized the organization of the human Apo2L gene and its promoter region. The Apo2L gene spans approximately 20 kb and is composed of five exons. The 1.2-kb Apo2L promoter region upstream of the translation initiation codon was cloned, its transcription start site defined, and several putative transcription factor binding sites identified. Luciferase reporter constructs were transfected into Jurkat cells and shown to be induced by IFNs. Deletion analysis indicates that the Apo2L promoter region between nucleotides 126 and 33 upstream of the transcriptional start site controls the expression of the Apo2L gene following IFN-beta treatment.