Abundant constitutive expression of the immediate-early 94K protein from cytomegalovirus (Colburn) in a DNA-transfected mouse cell line
A 94-kilodalton phosphoprotein known as IE94 is the only viral polypeptide synthesized in abundance under immediate-early conditions after infection by cytomegalovirus (CMV) strain Colburn in either permissive primate or nonpermissive rodent cells. The authors isolated a clonal Ltk/sup +/ cell line which expressed the /sup 35/methionine-labeled IE94 polypeptide in sufficient abundance to be visualized directly in autoradiographs after gel electrophoresis of total-cell-culture protein extracts. The IE94 polypeptide synthesized in the transfected cells was indistinguishable in size and overall net charge from that produced in virus-infected cells. In addition, the IE94 protein expressed in LH/sub 2/p198-3 cells was phosphorylated (presumably by a cellular protein kinase) and generated similar phosphopeptide patterns after partial tryptic digestion to those obtained with the CMV IE94 protein from infected cells. The cell line contained two to four stably integrated copies of the IE94 gene and synthesized a single virus-specific mRNA of 2.5 kilobases detectable on Northern blots. A new antigen, detectable by indirect anticomplement immunofluorescence with monoclonal antibody against the human CMV IE68 protein, was present in the nuclei of more than 95% of the LH/sub 2/l198-3 cells. This evidence suggests that (unlike most herpesvirus genes) the CMV IE94 gene, together with its complex promoter and spliced mRNA structure, may contain all of the regulatory elements necessary for strong constitutive expression in mammalian cells in the absence of other viral factors.
- Research Organization:
- Dept. of Pharmacology and Experimental Therapeutics, Johns Hopkins Univ. School of Medicine, Baltimore, MD 21205
- OSTI ID:
- 5942363
- Journal Information:
- Mol. Cell. Biol.; (United States), Journal Name: Mol. Cell. Biol.; (United States) Vol. 4:10; ISSN MCEBD
- Country of Publication:
- United States
- Language:
- English
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59 BASIC BIOLOGICAL SCIENCES
AMINO ACIDS
ANIMALS
ANTIBODIES
ANTIGENS
AROMATICS
AUTORADIOGRAPHY
AZAARENES
AZINES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOSYNTHESIS
CARBOXYLIC ACIDS
CELL CONSTITUENTS
CELL CULTURES
CELL TRANSFORMATIONS
CHEMICAL REACTIONS
CLONE CELLS
CULTURE MEDIA
DAYS LIVING RADIOISOTOPES
DECOMPOSITION
DISEASES
DNA
DRUGS
ELECTROPHORESIS
ENZYMATIC HYDROLYSIS
ENZYMES
EVEN-ODD NUCLEI
GENES
GENETIC MAPPING
GENOME MUTATIONS
HERPES SIMPLEX
HETEROCYCLIC COMPOUNDS
HYDROLYSIS
HYDROXY COMPOUNDS
HYPOXANTHINE
IMAGES
INFECTIOUS DISEASES
ISOTOPES
LABELLING
LIGHT NUCLEI
LIPOTROPIC FACTORS
LYSIS
MAMMALS
MAPPING
MESSENGER-RNA
METHIONINE
MOLECULAR BIOLOGY
MONOCLONAL ANTIBODIES
MUTATIONS
NUCLEI
NUCLEIC ACIDS
NUCLEOSIDES
NUCLEOTIDES
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
ORGANIC SULFUR COMPOUNDS
PHOSPHOPROTEINS
PHOSPHORUS-GROUP TRANSFERASES
PHOSPHOTRANSFERASES
PLASMIDS
PRECIPITATION
PROTEINS
PURINES
PYRIMIDINES
RADIOISOTOPES
RECOMBINANT DNA
RIBOSIDES
RNA
RODENTS
SEPARATION PROCESSES
SKIN DISEASES
SOLVOLYSIS
SPECIFICITY
STRAND BREAKS
SULFUR 35
SULFUR ISOTOPES
SYNTHESIS
THYMIDINE
TRANSFERASES
VERTEBRATES
VIRAL DISEASES