scholarly journals Cell-specific activity of the modulator region in the human cytomegalovirus major immediate-early gene.

1989 ◽  
Vol 9 (3) ◽  
pp. 1342-1345 ◽  
Author(s):  
H Lubon ◽  
P Ghazal ◽  
L Hennighausen ◽  
C Reynolds-Kohler ◽  
C Lockshin ◽  
...  
Keyword(s):  
Human Cytomegalovirus ◽  
Transcriptional Activation ◽  
Transcriptional Repression ◽  
Specific Activity ◽  
Gene Promoter ◽  
Early Gene ◽  
Immediate Early ◽  
Upstream Region ◽  
Early Promoter ◽  
Major Immediate Early Promoter

In this paper we demonstrate that modulator sequences upstream of the enhancer of the major immediate-early promoter of human cytomegalovirus exert a differential effect on the level of transcription in a variety of cells and that this region has the capacity to interact with specific nuclear protein. Depending on the cell type, these modulator sequences increased or decreased transcriptional activation from the IE1 gene promoter-enhancer. The cell lines identified in this report should be useful to study the molecular mechanism of cell-specific transcriptional repression and activation exerted by the major immediate-early promoter upstream region.

scholarly journals Cell-specific activity of the modulator region in the human cytomegalovirus major immediate-early gene

1989 ◽  
Vol 9 (3) ◽  
pp. 1342-1345
Author(s):  
H Lubon ◽  
P Ghazal ◽  
L Hennighausen ◽  
C Reynolds-Kohler ◽  
C Lockshin ◽  
...  
Keyword(s):  
Human Cytomegalovirus ◽  
Transcriptional Activation ◽  
Transcriptional Repression ◽  
Specific Activity ◽  
Gene Promoter ◽  
Early Gene ◽  
Immediate Early ◽  
Upstream Region ◽  
Early Promoter ◽  
Major Immediate Early Promoter

In this paper we demonstrate that modulator sequences upstream of the enhancer of the major immediate-early promoter of human cytomegalovirus exert a differential effect on the level of transcription in a variety of cells and that this region has the capacity to interact with specific nuclear protein. Depending on the cell type, these modulator sequences increased or decreased transcriptional activation from the IE1 gene promoter-enhancer. The cell lines identified in this report should be useful to study the molecular mechanism of cell-specific transcriptional repression and activation exerted by the major immediate-early promoter upstream region.

scholarly journals In vivo and in vitro analysis of transcriptional activation mediated by the human cytomegalovirus major immediate-early proteins.

1993 ◽  
Vol 13 (2) ◽  
pp. 1238-1250 ◽  
Author(s):  
K M Klucher ◽  
M Sommer ◽  
J T Kadonaga ◽  
D H Spector
Keyword(s):  
Human Cytomegalovirus ◽  
Hela Cells ◽  
Transcriptional Activation ◽  
Histone H1 ◽  
Early Gene ◽  
Immediate Early ◽  
Early Promoter ◽  
Drosophila Embryos

To define mechanistically how the human cytomegalovirus (HCMV) major immediate-early (IE) proteins induce early-gene transcription, the IE1 72-kDa protein, the IE2 55-kDa protein, and the IE2 86-kDa protein were analyzed for their ability to activate transcription from an HCMV early promoter in vivo and in vitro. In transient-expression assays in U373MG astrocytoma/glioblastoma and HeLa cells, only the IE2 86-kDa protein was able to activate the HCMV early promoter to high levels. In HeLa cells, the IE1 72-kDa protein was able to activate the promoter to a low but detectable level, and the level of promoter activity observed in response to the IE2 86-kDa protein was increased synergistically following cotransfection of the constructs expressing both IE proteins. To examine the interaction of the HCMV IE proteins with the RNA polymerase II transcription machinery, we assayed the ability of Escherichia coli-synthesized proteins to activate the HCMV early promoter in nuclear extracts prepared from U373MG cells, HeLa cells, and Drosophila embryos. The results of the in vitro experiments correlated well with those obtained in vivo. The basal activity of the promoter was minimal in both the HeLa and U373MG extracts but was stimulated 6- to 10-fold by the IE2 86-kDa protein. With a histone H1-deficient extract from Drosophila embryos, the HCMV early promoter was quite active and was stimulated two- to fourfold by the IE2 86-kDa protein. Addition of histone H1 at 1 molecule per 40 to 50 bp of DNA template significantly repressed basal transcription from this promoter. However, the IE2 86-kDa protein, but none of the other IE proteins, was able to counteract the H1-mediated repression and stimulate transcription at least 10- to 20-fold. The promoter specificity of the activation was demonstrated by the inability of the IE2 86-kDa protein to activate the Drosophila Krüppel promoter in either the presence or absence of histone H1. These results suggest that one mechanism of transcription activation by the IE2 86-kDa protein involves antirepression.

scholarly journals The Human Cytomegalovirus UL35 Gene Encodes Two Proteins with Different Functions

2002 ◽  
Vol 76 (5) ◽  
pp. 2460-2468 ◽  
Author(s):  
Yingguang Liu ◽  
Bonita J. Biegalke
Keyword(s):  
Human Cytomegalovirus ◽  
Complex Structure ◽  
Protein A ◽  
Immediate Early Gene ◽  
Early Gene ◽  
Immediate Early ◽  
Terminal Portion ◽  
Virion Protein ◽  
Early Promoter ◽  
Major Immediate Early Promoter

ABSTRACT The human cytomegalovirus (HCMV) virion is a complex structure that contains at least 30 proteins, many of which have been identified. We determined that the HCMV UL35 gene encodes two proteins, including a previously unidentified virion protein. A 22-kDa phosphoprotein (ppUL35A) was translated from a 1.2-kb UL35 transcript by 4 h postinfection; a second phosphoprotein of 75 kDa (ppUL35) was translated from a 2.2-kb transcript predominantly late in infection. The 22-kDa protein localized to the nucleus, while the 75-kDa protein localized to the juxtanuclear compartment and was packaged into virion particles. The 22-kDa protein was identical to the COOH-terminal end of the 75-kDa protein but was not found in virions, thus defining the NH2-terminal portion of the 75-kDa protein as essential for packaging. Expression of the 22-kDa protein inhibited activation of the major immediate-early promoter by ppUL82 (pp71), suggesting that the UL35 22-kDa protein may modulate expression of the major immediate-early gene.

scholarly journals Human cytomegalovirus major immediate early transcripts arise predominantly from the canonical major immediate early promoter in reactivating progenitor-derived dendritic cells

2020 ◽  
Vol 101 (6) ◽  
pp. 635-644 ◽  
Author(s):  
Rebecca Mason ◽  
Ian J. Groves ◽  
Mark R. Wills ◽  
John H. Sinclair ◽  
Matthew B. Reeves
Keyword(s):  
Gene Expression ◽  
Dendritic Cells ◽  
Human Cytomegalovirus ◽  
Virus Production ◽  
Early Gene ◽  
Immediate Early ◽  
Genome Replication ◽  
Early Promoter ◽  
Major Immediate Early Promoter

Human cytomegalovirus latency and reactivation is a major source of morbidity in immune-suppressed patient populations. Lifelong latent infections are established in CD34+progenitor cells in the bone marrow, which are hallmarked by a lack of major lytic gene expression, genome replication and virus production. A number of studies have shown that inhibition of the major immediate early promoter (MIEP) – the promoter that regulates immediate early (IE) gene expression – is important for the establishment of latency and that, by extension, reactivation requires reversal of this repression of the MIEP. The identification of novel promoters (termed ip1 and ip2) downstream of the MIEP that can drive IE gene expression has led to speculation over the precise role of the MIEP in reactivation. In this study we show that IE transcripts arise from both the MIEP and ip2 promoter in the THP1 cell macrophage cell line and also CD14+monocytes stimulated with phorbol ester. In contrast, we show that in in vitro generated dendritic cells or macrophages that support HCMV reactivation IE transcripts arise predominantly from the MIEP and not the intronic promoters. Furthermore, inhibition of histone modifying enzyme activity confirms the view that the MIEP is predominantly regulated by the activity of cellular chromatin. Finally, we observe that ip2-derived IE transcription is cycloheximide-sensitive in reactivating DCs, behaviour consistent with an early gene designation. Taken together, these data argue that MIEP activity is still important for HCMV reactivation but ip2 activity could play cell-type-specific roles in reactivation.

In vivo and in vitro analysis of transcriptional activation mediated by the human cytomegalovirus major immediate-early proteins

1993 ◽  
Vol 13 (2) ◽  
pp. 1238-1250
Author(s):  
K M Klucher ◽  
M Sommer ◽  
J T Kadonaga ◽  
D H Spector
Keyword(s):  
Human Cytomegalovirus ◽  
Hela Cells ◽  
Transcriptional Activation ◽  
Histone H1 ◽  
Early Gene ◽  
Immediate Early ◽  
Early Promoter ◽  
Drosophila Embryos

To define mechanistically how the human cytomegalovirus (HCMV) major immediate-early (IE) proteins induce early-gene transcription, the IE1 72-kDa protein, the IE2 55-kDa protein, and the IE2 86-kDa protein were analyzed for their ability to activate transcription from an HCMV early promoter in vivo and in vitro. In transient-expression assays in U373MG astrocytoma/glioblastoma and HeLa cells, only the IE2 86-kDa protein was able to activate the HCMV early promoter to high levels. In HeLa cells, the IE1 72-kDa protein was able to activate the promoter to a low but detectable level, and the level of promoter activity observed in response to the IE2 86-kDa protein was increased synergistically following cotransfection of the constructs expressing both IE proteins. To examine the interaction of the HCMV IE proteins with the RNA polymerase II transcription machinery, we assayed the ability of Escherichia coli-synthesized proteins to activate the HCMV early promoter in nuclear extracts prepared from U373MG cells, HeLa cells, and Drosophila embryos. The results of the in vitro experiments correlated well with those obtained in vivo. The basal activity of the promoter was minimal in both the HeLa and U373MG extracts but was stimulated 6- to 10-fold by the IE2 86-kDa protein. With a histone H1-deficient extract from Drosophila embryos, the HCMV early promoter was quite active and was stimulated two- to fourfold by the IE2 86-kDa protein. Addition of histone H1 at 1 molecule per 40 to 50 bp of DNA template significantly repressed basal transcription from this promoter. However, the IE2 86-kDa protein, but none of the other IE proteins, was able to counteract the H1-mediated repression and stimulate transcription at least 10- to 20-fold. The promoter specificity of the activation was demonstrated by the inability of the IE2 86-kDa protein to activate the Drosophila Krüppel promoter in either the presence or absence of histone H1. These results suggest that one mechanism of transcription activation by the IE2 86-kDa protein involves antirepression.

Correlation Between Expression of Recombinant Proteins and Abundance of H3K4Me3 on the Enhancer of Human Cytomegalovirus Major Immediate-Early Promoter

2017 ◽  
Vol 59 (8) ◽  
pp. 315-322 ◽  
Author(s):  
Benjamin P. C. Soo ◽  
Julian Tay ◽  
Shirelle Ng ◽  
Steven C. L. Ho ◽  
Yuansheng Yang ◽  
...  
Keyword(s):  
Recombinant Proteins ◽  
Human Cytomegalovirus ◽  
Immediate Early ◽  
Early Promoter ◽  
Major Immediate Early Promoter

Tissue and cell-specific transcriptional activity of the human cytomegalovirus immediate early gene promoter (UL123) in zebrafish

2013 ◽  
Vol 13 (3-4) ◽  
pp. 91-103 ◽  
Author(s):  
Vanessa Mella-Alvarado ◽  
Aude Gautier ◽  
Florence Le Gac ◽  
Jean-Jacques Lareyre
Keyword(s):  
Human Cytomegalovirus ◽  
Transcriptional Activity ◽  
Gene Promoter ◽  
Immediate Early Gene ◽  
Early Gene ◽  
Immediate Early

Interactions between cellular regulatory proteins and a unique sequence region in the human cytomegalovirus major immediate-early promoter

Virology ◽  
1990 ◽  
Vol 174 (1) ◽  
pp. 18-25 ◽  
Author(s):  
Peter Ghazal ◽  
Henryk Lubon ◽  
Catherine Reynolds-Kohler ◽  
Lothar Hennighausen ◽  
Jay A. Nelson
Keyword(s):  
Human Cytomegalovirus ◽  
Unique Sequence ◽  
Regulatory Proteins ◽  
Immediate Early ◽  
Early Promoter ◽  
Sequence Region ◽  
Major Immediate Early Promoter
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