Bovine leukemia virus (BLV) is the causative agent of bovine leukosis, a naturally occurring fatal disease in cattle. BLV transcription is regulated by cellular transcription factors and the virally encoded oncoprotein Tax. In this report, we investigated the functional role of the putative NF-kappa B binding site recently identified in the BLV promoter; Our studies indicate that the kappa B binding motif acts as a functional enhancer in the presence of the cellular NF-kappa B proteins. Furthermore, the kappa B site together with a single 21-bp repeat confers strong activation of BLV transcription in the presence the NF-kappa B proteins and lax. These results suggest that cellular NF-kappa B may he involved in the regulation of BLV transcription and activation of the virus from latency, (C) 1998 Academic Press.