The leader RNA transcript from the 3' end of the genome of vesicular stomatitis virus (VSV) has been detected in both the nucleus and cytoplasm of infected baby hamster kidney (BHK) cells. In the cytoplasm, leader RNA accumulated gradually throughout the infection to about 200 molecules per cell at 6 hr after infection. In the nucleus, however, there was a sharp and rapid increase in the concentration of leader RNA to approximately equal to 300 molecules per cell at about 2 hr after infection that decreased rapidly by 3 hr. This report presents evidence for nuclear localization of transcription products of a (-)-strand RNA virus other than influenza and supports the hypothesis that the leader RNA plays a role in the shutoff of host cell transcription.