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Title: Abundant constitutive expression of the immediate-early 94K protein from cytomegalovirus (Colburn) in a DNA-transfected mouse cell line

Abstract

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 andmore » 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.« less

Authors:
; ;
Publication Date:
Research Org.:
Dept. of Pharmacology and Experimental Therapeutics, Johns Hopkins Univ. School of Medicine, Baltimore, MD 21205
OSTI Identifier:
5942363
Resource Type:
Journal Article
Resource Relation:
Journal Name: Mol. Cell. Biol.; (United States); Journal Volume: 4:10
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; CELL TRANSFORMATIONS; MOLECULAR BIOLOGY; HERPES SIMPLEX; GENETIC MAPPING; MESSENGER-RNA; BIOSYNTHESIS; SPECIFICITY; PHOSPHOPROTEINS; ENZYMATIC HYDROLYSIS; GENES; ANTIGENS; AUTORADIOGRAPHY; CELL CULTURES; CLONE CELLS; CULTURE MEDIA; ELECTROPHORESIS; GENOME MUTATIONS; HYPOXANTHINE; IMAGES; LABELLING; METHIONINE; MONOCLONAL ANTIBODIES; PHOSPHOTRANSFERASES; PLASMIDS; PRECIPITATION; RECOMBINANT DNA; RODENTS; STRAND BREAKS; SULFUR 35; THYMIDINE; AMINO ACIDS; ANIMALS; ANTIBODIES; AROMATICS; AZAARENES; AZINES; BETA DECAY RADIOISOTOPES; BETA-MINUS DECAY RADIOISOTOPES; CARBOXYLIC ACIDS; CELL CONSTITUENTS; CHEMICAL REACTIONS; DAYS LIVING RADIOISOTOPES; DECOMPOSITION; DISEASES; DNA; DRUGS; ENZYMES; EVEN-ODD NUCLEI; HETEROCYCLIC COMPOUNDS; HYDROLYSIS; HYDROXY COMPOUNDS; INFECTIOUS DISEASES; ISOTOPES; LIGHT NUCLEI; LIPOTROPIC FACTORS; LYSIS; MAMMALS; MAPPING; MUTATIONS; NUCLEI; NUCLEIC ACIDS; NUCLEOSIDES; NUCLEOTIDES; ORGANIC ACIDS; ORGANIC COMPOUNDS; ORGANIC NITROGEN COMPOUNDS; ORGANIC SULFUR COMPOUNDS; PHOSPHORUS-GROUP TRANSFERASES; PROTEINS; PURINES; PYRIMIDINES; RADIOISOTOPES; RIBOSIDES; RNA; SEPARATION PROCESSES; SKIN DISEASES; SOLVOLYSIS; SULFUR ISOTOPES; SYNTHESIS; TRANSFERASES; VERTEBRATES; VIRAL DISEASES; 550201* - Biochemistry- Tracer Techniques; 550401 - Genetics- Tracer Techniques

Citation Formats

Jeang, K.T., Cho, M.S., and Hayward, G.S. Abundant constitutive expression of the immediate-early 94K protein from cytomegalovirus (Colburn) in a DNA-transfected mouse cell line. United States: N. p., 1984. Web. doi:10.1128/MCB.4.10.2214.
Jeang, K.T., Cho, M.S., & Hayward, G.S. Abundant constitutive expression of the immediate-early 94K protein from cytomegalovirus (Colburn) in a DNA-transfected mouse cell line. United States. doi:10.1128/MCB.4.10.2214.
Jeang, K.T., Cho, M.S., and Hayward, G.S. Mon . "Abundant constitutive expression of the immediate-early 94K protein from cytomegalovirus (Colburn) in a DNA-transfected mouse cell line". United States. doi:10.1128/MCB.4.10.2214.
@article{osti_5942363,
title = {Abundant constitutive expression of the immediate-early 94K protein from cytomegalovirus (Colburn) in a DNA-transfected mouse cell line},
author = {Jeang, K.T. and Cho, M.S. and Hayward, G.S.},
abstractNote = {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.},
doi = {10.1128/MCB.4.10.2214},
journal = {Mol. Cell. Biol.; (United States)},
number = ,
volume = 4:10,
place = {United States},
year = {Mon Oct 01 00:00:00 EDT 1984},
month = {Mon Oct 01 00:00:00 EDT 1984}
}