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  • American Society for Microbiology  (5)
  • Ghazal, P  (5)
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  • American Society for Microbiology  (5)
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  • 1
    Online-Ressource
    Online-Ressource
    Developmental analysis of the cytomegalovirus enhancer in transgenic animals
    American Society for Microbiology ; 1996
    In:  Journal of Virology Vol. 70, No. 5 ( 1996-05), p. 3215-3226
    Details
    In: Journal of Virology, American Society for Microbiology, Vol. 70, No. 5 ( 1996-05), p. 3215-3226
    Kurzfassung: The major immediate-early promoter (MIEP) of human, cytomegalovirus (HCMV) constitutes a primary genetic switch for viral activation. In this study, regulation of the enhancer-containing segment (nucleotides -670 to +54) of the HCMV MIEP attached to the 1acZ reporter gene was examined in the developing embryos of transgenic mice to identify temporal and tissue-specific expression. We find that the transgene reporter is first detected as a dorsal stripe of expression in the neural folds of embryos at day 8.5 postcoitum (p.c.). A broad expression pattern is exhibited in embryos at day 9.5 p.c. This pattern becomes more restricted by day 10.5 p.c. as organogenesis progresses. By day 14.5 p.c., prominent expression is observed in a subpopulation of central nervous system cells and spinal ganglia, endothelial cells, muscle, skin, thyroid, parathyroid, kidney, lung, liver, and gut cells, and the pancreas and submandibular and pituitary glands. This distribution pattern is discussed in relation to human congenital HCMV infection. These results suggest that the transcriptional activity of the HCMV MIEP may determine in part, the ability of the virus to specifically target developing fetal tissues in utero.
    Materialart: Online-Ressource
    ISSN: 0022-538X , 1098-5514
    URL: Article
    DOI: 10.1128/jvi.70.5.3215-3226.1996
    Sprache: Englisch
    Verlag: American Society for Microbiology
    Publikationsdatum: 1996
    ZDB Id: 1495529-5
    Standort Signatur Einschränkungen Verfügbarkeit
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    Online-Ressource
  • 2
    Online-Ressource
    Online-Ressource
    The enhancer domain of the human cytomegalovirus major immediate-early promoter determines cell type-specific expression in transgenic mice
    American Society for Microbiology ; 1996
    In:  Journal of Virology Vol. 70, No. 5 ( 1996-05), p. 3207-3214
    Details
    In: Journal of Virology, American Society for Microbiology, Vol. 70, No. 5 ( 1996-05), p. 3207-3214
    Kurzfassung: The human cytomegalovirus (HCMV) major immediate-early promoter (MIEP) is one of the first promoters to activate upon infection. To examine HCMV MIEP tissue-specific expression, transgenic mice were established containing the lacZ gene regulated by the MIEP (nucleotides -670 to +54). In the transgenic mice, lacZ expression was demonstrated in 19 of 29 tissues tested by histochemical and immunochemical analyses. These tissues included brain, eye, spinal cord, esophagus, stomach, pancreas, kidney, bladder, testis, ovary, spleen, salivary gland, thymus, bone marrow, skin, cartilage, and cardiac, striated and smooth muscles. Although expression was observed in multiple organs, promoter activity was restricted to specific cell types. The cell types which demonstrated HCMV MIEP expression included retinal cells of the eye, ductile cells of the salivary gland, exocrine cells of the pancreas, mucosal cells of the stomach and intestine, neuronal cells of the brain, muscle fibers, thecal cells of the corpus luteum, and Leydig and sperm cells of the testis. These observations indicate that the HCMV MIEP is not a pan-specific promoter and that the majority of expressing tissues correlate with tissues naturally infected by the virus in the human host.
    Materialart: Online-Ressource
    ISSN: 0022-538X , 1098-5514
    URL: Article
    DOI: 10.1128/jvi.70.5.3207-3214.1996
    Sprache: Englisch
    Verlag: American Society for Microbiology
    Publikationsdatum: 1996
    ZDB Id: 1495529-5
    Standort Signatur Einschränkungen Verfügbarkeit
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    Online-Ressource
  • 3
    Online-Ressource
    Online-Ressource
    Direct interaction of the human cytomegalovirus IE86 protein with the cis repression signal does not preclude TBP from binding to the TATA box
    American Society for Microbiology ; 1993
    In:  Journal of Virology Vol. 67, No. 9 ( 1993-09), p. 5595-5604
    Details
    In: Journal of Virology, American Society for Microbiology, Vol. 67, No. 9 ( 1993-09), p. 5595-5604
    Kurzfassung: The human cytomegalovirus major immediate-early gene encodes several protein isoforms which autoregulate the major immediate-early promoter (MIEP). One of these isoforms, the IE86 protein, represses the MIEP through a DNA sequence located between the TATA box and the transcription initiation site, designated the cis repression signal (crs). Through mutational analysis, amino acid domains within IE86 responsible for binding the crs element were located at the C terminus. Mutation of the putative zinc finger domain, which precluded IE86 from binding DNA, converted the protein from a repressor of MIEP transcription into an activator. DNase I protection analysis demonstrated that the IE86 footprint overlapped the sequence protected by the TATA-binding protein (TBP). Investigation of whether IE86 was able to displace TBP from DNA revealed that both proteins could bind DNA simultaneously. However, higher concentrations of IE86 were required to obtain protection of the crs element in the presence of prebound TBP. Similarly, higher concentrations of TBP were required to obtain protection in the presence of prebound IE86. These observations indicate that steric hinderance impairs but does not prevent both proteins from binding DNA synchronously.
    Materialart: Online-Ressource
    ISSN: 0022-538X , 1098-5514
    URL: Article
    DOI: 10.1128/jvi.67.9.5595-5604.1993
    Sprache: Englisch
    Verlag: American Society for Microbiology
    Publikationsdatum: 1993
    ZDB Id: 1495529-5
    Standort Signatur Einschränkungen Verfügbarkeit
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    Online-Ressource
  • 4
    Online-Ressource
    Online-Ressource
    Human cytomegalovirus IE86 protein interacts with promoter-bound TATA-binding protein via a specific region distinct from the autorepression domain
    American Society for Microbiology ; 1993
    In:  Journal of Virology Vol. 67, No. 12 ( 1993-12), p. 7539-7546
    Details
    In: Journal of Virology, American Society for Microbiology, Vol. 67, No. 12 ( 1993-12), p. 7539-7546
    Kurzfassung: The major immediate-early gene of human cytomegalovirus encodes several isoforms of an immediate-early protein which has distinct transcriptional regulatory properties. The IE86 isoform autorepresses the major immediate-early promoter by directly binding the cis repression signal element located between the TATA box and the mRNA cap site. In addition to this activity, IE86 stimulates other viral and cellular promoters. One mechanism by which eukaryotic regulatory proteins are thought to stimulate transcription is by contacting one or more general transcription factors. We show that the IE86 protein physically interacts with the DNA-binding subunit (TATA-binding protein) human transcription factor IID via the TATA-binding protein-contacting domain in the N terminus of IE86. In a mobility shift assay, IE86 was also observed to stabilize the binding of TATA-binding protein to promoter DNA. The domains within IE86 responsible for mediating transactivation and repression functioned independently. These experiments thus demonstrate the elegant ability of human cytomegalovirus to join different protein domains to produce distinct multifunctional proteins.
    Materialart: Online-Ressource
    ISSN: 0022-538X , 1098-5514
    URL: Article
    DOI: 10.1128/jvi.67.12.7539-7546.1993
    Sprache: Englisch
    Verlag: American Society for Microbiology
    Publikationsdatum: 1993
    ZDB Id: 1495529-5
    Standort Signatur Einschränkungen Verfügbarkeit
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    Online-Ressource
  • 5
    Online-Ressource
    Online-Ressource
    Promoter-specific trans activation and repression by human cytomegalovirus immediate-early proteins involves common and unique protein domains
    American Society for Microbiology ; 1990
    In:  Journal of Virology Vol. 64, No. 4 ( 1990-04), p. 1556-1565
    Details
    In: Journal of Virology, American Society for Microbiology, Vol. 64, No. 4 ( 1990-04), p. 1556-1565
    Kurzfassung: trans activation of promoters by viral regulatory proteins provides a useful tool to study coordinate control of gene expression. Immediate-early (IE) regions 1 and 2 of human cytomegalovirus (CMV) code for a series of proteins that originate from differentially spliced mRNAs. These IE proteins are proposed to regulate the temporal expression of the viral genome. To examine the structure and function of the IE proteins, we used linker insertion mutagenesis of the IE gene region as well as cDNA expression vector cloning of the abundant IE mRNAs. We showed that IE1 and IE2 proteins of CMV exhibit promoter-specific differences in their modes of action by either trans activating early and IE promoters or repressing the major IE promoter (MIEP). Transient cotransfection experiments with permissive human cells revealed a synergistic interaction between the 72- and the 86-kilodalton (kDa) IE proteins in trans activating an early promoter. In addition, transfection studies revealed that the 72-kDa protein was capable of trans activating the MIEP. In contrast, the 86-kDa protein specifically repressed the MIEP and this repression was suppressed by the 72-kDa protein. Furthermore, observations based on the primary sequence structure revealed a modular arrangement of putative regulatory motifs that could either potentiate or repress gene expression. These modular domains are either shared or unique among the IE proteins. From these data, we propose a model for IE protein function in the coordinate control of CMV gene expression.
    Materialart: Online-Ressource
    ISSN: 0022-538X , 1098-5514
    URL: Article
    DOI: 10.1128/jvi.64.4.1556-1565.1990
    Sprache: Englisch
    Verlag: American Society for Microbiology
    Publikationsdatum: 1990
    ZDB Id: 1495529-5
    Standort Signatur Einschränkungen Verfügbarkeit
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    Online-Ressource
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