Differential Expression of the Major Immediate Early Gene of Human Cytomegalovirus

Abstract

We prepared a murine monoclonal antibody reactive to a human cytomegalovirus (HCMV)-induced nuclear protein with an Mr of 68000. Expression of the 68K protein was compared with the major immediate early (IE) 72K protein in various cell types after infection with HCMV or microinjection of plasmid DNA containing the major IE gene. The 68K nuclear protein was detected 2 to 3 h after appearance of the 72K protein in human embryonal lung (HEL) cells infected with HCMV. The 68K protein was distributed throughout the cytoplasm in the late phase of infection, while the 72K protein remained chiefly in the nucleus. The 68K protein was barely detected in the cells under IE conditions by immunoprecipitation, but, together with the 72K protein, it was expressed after microinjection of cloned DNA, containing only the major IE region (region 1), into the nuclei of HEL cells. The 72K protein was expressed in nuclei 2 h after microinjection, whereas the 68K protein was detected 4 to 5 h after the injection. The 68K protein was expressed after microinjection in non-permissive rodent fibroblasts or non-permissive transformed human cells in which these proteins were not expressed after viral infection.Immunoprecipitations after chase-labelling from IE conditions or after partial digestions suggested that the 68K protein is neither a degradation nor a modification product of the major IE 72K protein.

Later Passages of Neural Progenitor Cells from Neonatal Brain Are More Permissive for Human Cytomegalovirus Infection

“…3 to 6 and 8). These results are consistent with results described previously (40,42,43,45,63,64). No notable differences in infection were observed among the same-passage NPCs originating from different gestational ages (Fig.…”

Subcellular distribution of the major immediate early proteins of human cytomegalovirus changes during infection

“…We have reported already that the 68K protein recognized by monoclonal antibody (MAb) e-8 is expressed from IE1, which also encodes the major IE protein of 72K (Tsutsui & Nogami-Satake, 1990). In the present study, we show that the ultrastructural localization of the 72K and 68K proteins in permissive cells changes during the course of infection.…”

“…Virus stocks were prepared in human embryonic lung (HEL) cells and grown in Dulbecco's modified Eagle's medium containing 10% foetal calf serum, penicillin (100 units/ml) and streptomycin (50~tg/ml). Preparation of MAb e-8, which reacts with the 68K protein of HCMV-infected cells, has been described previously (Tsutsui & Nogami-Satake, 1990). The 68K protein detected by MAb e-8 is expressed after microinjection of ctoned DNA containing only the major IE region (/El) of HCMV (Tsutsui & Nogami-Satake, 1990).…”

Relationship between HOX2 homeobox gene expression and the human cytomegalovirus immediate early genes

“…After incubation with antidigoxigenin antibody conjugated to alkaline phosphatase, the slides were visualized according to the manufacturer's instruction (Boehringer Mannheim). After in situ hybridization, the slides were subjected to immunofluorescence using the monoclonal antibody E3, which is specific for the 72K IE protein of HCMV (provided by L. C. Goldstein, Genetic System Corporation, Seattle, U.S.A.) (Goldstein et al, 1982) as described previously (Tsutsui & Nogami-Satake, 1990).…”

“…HCMV strain AD169 was obtained from Dr Y. Nishiyama, Nagoya University School of Medicine, 0001-0557 © 1992 SGM Japan. Virus stocks were prepared in human embryonic lung (HEL) cells and grown in Dulbecco's modified Eagle's medium (DMEM) containing 10~ foetal calf serum (FCS), penicillin (100 units/ml) and streptomycin (50 p.g/ml) (Tsutsui & Nogami-Satake, 1990). Virus was quantified by the plaque assay method of Wentworth & French (1970).…”