Previously, we have shown that bovine herpesvirus 1 (BHV-1) down-regulates the expression of major histocompatibility complex class I molecules by interfering with transport of peptides by the transporter associated with antigen processing (TAP). Further studies revealed that BHV-1 down-regulates the expression of mRNA for class I molecules and other cellular proteins. To further elucidate the mechanisms of down-regulation of class I molecules, a virion host shut-off (vhs) deletion mutant was generated. The mutant, like the wildtype (wt) virus, interfered with transport of peptides by the TAP, and down-regulated cell surface expression of class I molecules. However, unlike the wt virus, the mutant did not impair the synthesis of class I molecules. These results indicate that down-regulation of class I molecules by BHV-1 is mediated by vhs activity of the virus, as well as mechanisms specifically directed at the class I pathway. Absence of vhs activity should result in decreased pathogenicity and enhanced immunogenicity of BHV-1 vhs deletion mutant, making it a better vaccine candidate.