Constitutive and retinoic acid-inducible expression of cytomegalovirus immediate-early genes in human teratocarcinoma cells

Human teratocarcinoma stem cells are nonpermissive for human cytomegalovirus (HCMV) but become permissive after being induced to differentiate by treatment with retinoic acid. We show that in uninduced teratocarcinoma stem cells, and also in transformed human 293 cells expressing adenovirus E1a gene products, the HCMV immediate-early (IE) 68,000-molecular-weight polypeptide (68K polypeptide) was not expressed, and consequently input viral genomes were not replicated. However, after differentiation of the teratocarcinoma cells, synthesis of the HCMV IE 68K polypeptide was induced, and viral DNA replication occurred. In contrast to our observations for HCMV, simian cytomegalovirus (SCMV) displayed constitutive expression of its analogous IE 94K polypeptide, and the input SCMV genomes were replicated in both uninduced stem cells and 293 cells. Since little, if any, HCMV IE RNA was detectable in human teratocarcinoma or 293 cells after infection under IE conditions, we suggest that a direct transcriptional block to permissivity occurs in these cells. The presence of tandemly repeated sequences which bind nuclear factor I protein in the promoter for the SCMV IE 94K polypeptide gene but not in the promoter for the HCMV IE 68K polypeptide gene may allow the expression of the simian but not of the human IE gene product in transformed cells.

[1]  J. Bishop,et al.  The protein products of the myc and myb oncogenes and adenovirus E1a are structurally related , 1983, Nature.

[2]  W. Rutter,et al.  Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. , 1979, Biochemistry.

[3]  P. Thomas,et al.  Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. , 1980, Proceedings of the National Academy of Sciences of the United States of America.

[4]  J. Demarchi Nature of the block in the expression of some early virus genes in cells abortively infected with human cytomegalovirus. , 1983, Virology.

[5]  D. R. Thomsen,et al.  Structural analysis of the major immediate early gene of human cytomegalovirus , 1984, Journal of virology.

[6]  P. Reitsma,et al.  Regulation of myc gene expression in HL-60 leukaemia cells by a vitamin D metabolite , 1983, Nature.

[7]  R. Huebner,et al.  Cytopathogenic agent resembling human salivary gland virus recovered from tissue cultures of human adenoids. , 1956, Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine.

[8]  T. Curran,et al.  Induction of c-fos during myelomonocytic differentiation and macrophage proliferation , 1985, Nature.

[9]  D. R. Thomsen,et al.  Promoter-regulatory region of the major immediate early gene of human cytomegalovirus. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[10]  P. Chambon,et al.  Adenovirus-2 E1A products repress enhancer-induced stimulation of transcription , 1984, Nature.

[11]  B. Fleckenstein,et al.  Cloning of the complete human cytomegalovirus genome in cosmids. , 1982, Gene.

[12]  D. Spector,et al.  Cleavage maps for human cytomegalovirus DNA strain AD169 for restriction endonucleases EcoRI, BglII, and HindIII , 1982, Journal of virology.

[13]  Michael Boshart,et al.  A very strong enhancer is located upstream of an immediate early gene of human cytomegalovirus , 1985, Cell.

[14]  T. H. Weller,et al.  Isolation of Intranuclear Inclusion Producing Agents from Infants with Illnesses Resembling Cytomegalic Inclusion Disease.∗ † , 1957, Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine.

[15]  M. Stinski,et al.  Activation of the major immediate early gene of human cytomegalovirus by cis-acting elements in the promoter-regulatory sequence and by virus-specific trans-acting components , 1985, Journal of virology.

[16]  M. Stinski Sequence of protein synthesis in cells infected by human cytomegalovirus: early and late virus-induced polypeptides , 1978, Journal of virology.

[17]  M. Oldstone,et al.  Cytomegalovirus causes a latent infection in undifferentiated cells and is activated by induction of cell differentiation , 1981, The Journal of experimental medicine.

[18]  K. Jeang,et al.  A cycloheximide-enhanced protein in cytomegalovirus-infected cells. , 1980, Virology.

[19]  R. Wides,et al.  Structure and function of the adenovirus origin of replication , 1984, Cell.

[20]  J. Geelen,et al.  Human cytomegalovirus DNA. I. Molecular weight and infectivity , 1978, Journal of virology.

[21]  Margaret G. Smith Propagation in Tissue Cultures of a Cytopathogenic Virus from Human Salivary Gland Virus (SGV) Disease.∗ , 1956, Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine.

[22]  D. R. Thomsen,et al.  DNA of human cytomegalovirus: size heterogeneity and defectiveness resulting from serial undiluted passage , 1979, Journal of virology.

[23]  G. Hayward,et al.  Replicative forms of human cytomegalovirus DNA with joined termini are found in permissively infected human cells but not in non-permissive Balb/c-3T3 mouse cells. , 1983, The Journal of general virology.

[24]  K. Jeang,et al.  Characterization of cytomegalovirus immediate-early genes. I. Nonpermissive rodent cells overproduce the IE94K protein form CMV (Colburn). , 1982, Virology.

[25]  J. Demarchi Human cytomegalovirus DNA: restriction enzyme cleavage maps and map locations for immediate-early, early, and late RNAs. , 1981, Virology.

[26]  G. Khoury,et al.  Differentiation as a requirement for simian virus 40 gene expression in F-9 embryonal carcinoma cells. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[27]  G. Hayward,et al.  STRUCTURAL ORGANIZATION OF THE DNA MOLECULES FROM HUMAN CYTOMEGALOVIRUS , 1980 .

[28]  P. Andrews Retinoic acid induces neuronal differentiation of a cloned human embryonal carcinoma cell line in vitro. , 1984, Developmental biology.

[29]  P. Andrews,et al.  Cytomegalovirus replicates in differentiated but not in undifferentiated human embryonal carcinoma cells. , 1984, Science.

[30]  W. Gibson Immediate-early proteins of human cytomegalovirus strains AD 169, Davis, and Towne differ in electrophoretic mobility. , 1981, Virology.

[31]  Robert A. Weinberg,et al.  Tumorigenic conversion of primary embryo fibroblasts requires at least two cooperating oncogenes , 1983, Nature.

[32]  N C Dracopoli,et al.  Pluripotent embryonal carcinoma clones derived from the human teratocarcinoma cell line Tera-2. Differentiation in vivo and in vitro. , 1984, Laboratory investigation; a journal of technical methods and pathology.

[33]  E. Huang,et al.  Human Cytomegalovirus Genome: Partial Denaturation Map and Organization of Genome Sequences , 1977, Journal of virology.

[34]  F. Graham,et al.  Characteristics of a human cell line transformed by DNA from human adenovirus type 5. , 1977, The Journal of general virology.

[35]  P. Andrews,et al.  Cytomegalovirus infection of human teratocarcinoma cells in culture. , 1985, The Journal of general virology.

[36]  J. Nevins,et al.  Common control of the heat shock gene and early adenovirus genes: evidence for a cellular E1A-like activity , 1984, Molecular and cellular biology.