Gene expression in cells infected with gammaherpesvirus saimiri: properties of transcripts from two immediate-early genes

Abstract

During productive infections of cells with the gammaherpesvirus, herpesvirus saimiri (HVS), a polyadenylated RNA of 2.2-2.4 kb accumulates to form a large fraction of virus-specified RNA. This transcript is from the virus thymidylate synthase (TS) gene and its synthesis, like that of late mRNAs encoding the virus structural proteins, is sensitive to an inhibitor of virus DNA synthesis (phosphonoacetic acid, PAA). Transcription which is insensitive to PAA occurs from many parts of the HVS genome, including the EcoRI-D, EcoRI-E, EcoRI-I, and HindIII-G fragments. A 1.6-kb RNA from EcoRI-I/E and a 1.3-kb RNA from HindIII-G accumulate in HVS-infected cells incubated in the continuous presence of cycloheximide, and thus represent immediate-early (IE) class transcripts. The 1.3-kb message from HindIII-G is the predominant stable RNA under these conditions; accumulation of the 1.6-kb transcript from EcoRI-I/E (which encodes the previously characterized 52-kDa IE phosphoprotein) is markedly more dependent on the multiplicity of infection. The sequence of a 2.5-kbp region of the HindIII-G fragment has been determined and a single major open reading frame is present within the boundaries of the 1.3-kb IE RNA. Comparison of the amino acid sequence of the encoded protein (IE-G) with current databases of protein sequences failed to demonstrate significant similarities with herpesvirus proteins, but did detect a significant similarity with a region of the protein specified by an open reading frame in the LTR of mouse mammary tumor virus. The function of the IE gene in HindIII-G and the basis for the distinctive multiplicity dependence of IE transcription from the 52-kDa gene remain to be established.