Structural analysis of the major immediate early gene of human cytomegalovirus

Stenberg, R M; Thomsen, Darrell R.; Stinski, Mark F.

doi:10.1128/jvi.49.1.190-199.1984

Cited by 329 publications

(206 citation statements)

References 37 publications

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“…In the present study, we have confirmed their finding and The cloned HindIII C fragment of HCMV (Towne) DNA (6) was digested with EcoRI, and the EcoRI H fragment carrying the major immediate early (MIE) and immediate early 2 (IE2) genes (15)(16)(17)19) was subsequently cloned by being ligated into the EcoRI site of pSV2neo (13). This plasmid, designated pSV2neoEcoH ( Fig.…”

supporting

confidence: 76%

Expression of Human Cytomegalovirus Immediate Early Antigens Is Responsible for a Novel Chromatin Structure of Infected Human Embryonic Lung Cells

Maeda-Takekoshi

Takekoshi

Ihara

et al. 1988

Microbiology and Immunology

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supporting

confidence: 76%

Expression of Human Cytomegalovirus Immediate Early Antigens Is Responsible for a Novel Chromatin Structure of Infected Human Embryonic Lung Cells

Maeda-Takekoshi

Takekoshi

Ihara

et al. 1988

Microbiology and Immunology

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“…Genomic DNAs from the indicated clones were cleaved with DraI and blots hybridized with the cloned 300-400 bp target sequences as probes (Ti, T3 and T7, respectively). Retroposition results in new alleles (bands indicated by triangles), with increased sizes as compared with the unrearranged alleles (see arrows for the poly(A) tract of the reporter gene mRNA (Hegeman and Fiers, 1978); (ii) loss of the CMV promoter sequence, with the end point at the expected position for the transcription start site of the reporter gene mRNA for clones A-I and A-3 [within one nucleotide (Stenberg et al, 1984)], and 165 nucleotides downstream of the start site for clone A-7; and (iii) the presence, at both ends of the retroposed copies, of target site duplications of varying length (15 bp for clone A-1, 10 bp for clone A-3, no duplication for clone A-7, see Figure 3 legend). The structure of the retroposed mRNAs and flanking integration domains was analysed further by Southern blot, using the identified target sites as a probe ( Figure 3B).…”

Section: Resultsmentioning

confidence: 99%

mRNA retroposition in human cells: processed pseudogene formation.

et al. 1995

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'Corresponding authorUsing a sensitive assay for detection of reverse transcription events, we demonstrate that human HeLa cells can 'retropose', i.e. reverse transcribe and integrate, the mRNA of a naive reporter gene, at a low but detectable frequency. Furthermore, we show that the retroposed copies have all the hallmarks of the processed pseudogenes naturally found in the mammalian genome: they lack intron and 5' promoter sequence, they have acquired a 3' poly(A) tail, and they are flanked by short repeats (<15 bp) of target DNA sequence. These results demonstrate that human cells possess an endogenous reverse transcription activity, which is not restricted to transcripts of transposable elements, and which is likely to be involved in the formation, still ongoing, of a large fraction of the eukaryotic genome.

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“…The iel gene product is a nuclear phosphoprotein (Michelson-Fiske et al, 1977;Tanaka et al, 1979;Gibson, 1983;Stenberg et al, 1984). Although iel has not been shown to bind DNA directly, it has been recently reported to associate with metaphase chromosomes (La Femina et al, 1989).…”

Section: Discussionmentioning

confidence: 99%

NF-kappa B activation of the cytomegalovirus enhancer is mediated by a viral transactivator and by T cell stimulation.

et al. 1989

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The expression of cytomegalovirus alpha (immediate early) genes is under control of an enhancer that carries signals for strong constitutive expression as well as response elements for transactivation by viral proteins. We have used synthetic oligonucleotides representing the 16, 18 and 19 bp repeat elements within the enhancer to investigate the role of virus‐induced cellular transcription factors in enhancer activation. We show that the transcription factor NF‐kappa B, which binds to the 18 bp repeat, plays a central role in enhancer activation in infected human fibroblasts and that activation is mediated by the product of the viral gene ie1. The simian immunodeficiency virus kappa B site can functionally substitute for the 18 bp element in transient transactivation assays and can also compete efficiently for specific binding to the 18 bp repeat element in vitro. Point mutations in the NF‐kappa B site within the 18 bp element disrupt ie1‐mediated transactivation and binding. We have found that the characteristics of the 18 bp binding factor from human fibroblasts are indistinguishable from NF‐kappa B induced by phorbol ester plus mitogen treatment of T lymphocytes, as determined by gel mobility shift assay as well as protection of the binding site from chemical cleavage. Furthermore, T cell stimulation mediates activation of the viral enhancer via kappa B sites, an observation that may be important in the interaction of cytomegalovirus with the naturally infected human host. Thus, NF‐kappa B plays a central role as a target for enhancer activation via viral and cellular factors.

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Structural analysis of the major immediate early gene of human cytomegalovirus

Cited by 329 publications

References 37 publications

Expression of Human Cytomegalovirus Immediate Early Antigens Is Responsible for a Novel Chromatin Structure of Infected Human Embryonic Lung Cells

Expression of Human Cytomegalovirus Immediate Early Antigens Is Responsible for a Novel Chromatin Structure of Infected Human Embryonic Lung Cells

mRNA retroposition in human cells: processed pseudogene formation.

NF-kappa B activation of the cytomegalovirus enhancer is mediated by a viral transactivator and by T cell stimulation.

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