US22 gene family members m142 and m143 are essential for replication of murine cytomegalovirus (MCMV). Their transcripts are produced with immediate-early kinetics, but little else is known about these viral genes. Unlike their transcripts, the m142 and m143 gene products (pm142, pm143) were not expressed until early times post-infection, with levels increasing over the course of infection. Both pm142 and pm143 were predominantly cytoplasmic, but cellular fractionation studies confirmed that the proteins were present in the nucleus as well. In addition, pm142 was detected within the virion. Both the m142 and m143 promoters were strongly upregulated by viral infection or by MCMV IE1. However, UV-inactivated virus and IE3 upregulated only the m142 promoter. When tested for transcriptional transactivating activity, neither m142 nor m143 demonstrated significant activity, either alone or in combination with the major immediate-early gene products. This failure to transactivate, along with their essential nature, makes m142 and m143 unique among the immediate-early genes of the US22 gene family.