The ICP0 protein of herpes simplex virus type 1 (HSV-1) is a nuclear protein that possesses a well-characterized E3 ubiquitin ligase activity. This activity is responsible for the proteasomal-dependent degradation of several cellular proteins. This study shows that ICP0 induces the proteasomal-dependent degradation of the centromeric protein CENP-B in infected as well as ICP0-expressing cells. It is also shown that the ICP0-induced CENP-B degradation occurs as efficiently in human and mouse cells. CENP-B is one of the major proteins of centromeres and its degradation is likely to contribute to the severe damage induced to centromeres by ICP0.