Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:2.3.1.28 (chloramphenicol acetyltransferase)
5,100 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The sea urchin early H2A histone gene, like the other four members of the repeating units, is transiently expressed during very early development. To investigate the mechanisms underlying the faithful expression of the early H2A gene, we focused our attention on the modulator element. We showed by DNase I cleavage protection patterns that the modulator includes the upstream sequence element 1 (USE1) and mapped at nucleotides -137 to -108 in the early H2A gene promoter. Functional tests conducted by microinjection into sea urchin embryos then showed that the modulator element binds the transcriptional factor called modulator-binding factor 1 (MBF-1). We found in fact that coinjection of an excess of the MBF-1-binding site, either as the modulator or as the USE1, efficiently impaired the activity of the H2A promoter. An unexpected finding was the expression of the reporter gene from the early H2A promoter at the gastrula stage of embryonic development, when the early histone genes are transcriptionally silent. In addition, we also found that the modulator element was active at the gastrula stage. The potential enhancer activity of the modulator was tested by microinjecting several constructs containing single or multiple copies of the modulator element placed 5' or 3' to a thymidine kinase gene (tk) promoter in both sea urchin embryos and Xenopus laevis oocytes and determining the expression of a reporter chloramphenicol acetyltransferase gene under the control of the linked tk promoter. We found that an oligonucleotide bearing the MBF-1-binding site activates the expression of the reporter gene independently of the position and orientation. We conclude that the modulator binds the MBF-1 activator and that it is a transcriptional enhancer of the early H2A histone gene.
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PMID:Modulator factor-binding sequence of the sea urchin early histone H2A promoter acts as an enhancer element. 799 25

We have developed a novel, highly efficient DNA delivery system to accomplish gene transfer through the asialoglycoprotein receptor-mediated endocytosis pathway. Natural nuclear DNA-binding proteins, the histones (H1, H2a, H2b, H3, and H4), were modified and used as receptor-targeted DNA carriers. Galactosylated with a coupling agent, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, the histones and albumin were conjugated to DNA and then used to transfect HepG2 cells, which display the asialoglycoprotein receptor. The extent of galactosylation was determined for all histone subgroups and albumin with 14C-labeled galactose. A reporter gene for the bacterial chloramphenicol acetyltransferase (CAT), under the control of the 5' long terminal repeat (LTR) of Rous sarcoma virus, was used for comparisons of transfection efficiency of various carrier proteins. The CAT activity resulting from histone H1-mediated transfection was 1.66 unit per 10(6) cells, the highest among histone subgroups. The galactyosylated histone H1 was also eleven times more effective than the asialo-orosomucoid-polylysine. Ten galactosyl units are attached to histone H1 by the galactosylation reaction. Differences in the extent of galactosylation could not explain different transfection efficiencies among various proteins studied in this report. Treatment with galactose oxidase abolished the transfection ability of both the galactosylated histone H1 and asialo-orosomucoid. The intrinsic DNA-binding domains and nuclear location signal sequences are unique to histones as receptor-targeted DNA carriers, and are advantageous for effective gene delivery.
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PMID:Galactosylated histone-mediated gene transfer and expression. 804 1

We have identified a nuclear factor expressed in pro-B-, pre-B-, and B-cell lines that binds to two sites within the murine immunoglobulin heavy-chain (IgH) 3' alpha enhancer (3' alpha E). These sites were defined by oligonucleotide competition in an electrophoretic mobility shift assay (EMSA) and methylation interference footprinting. The 3' alpha E-binding factor is indistinguishable from NF-HB (B-lineage-specific nuclear factor that binds to the IgH gene) and the B-lineage-specific transcription factor BSAP by several criteria, including similar cell type distribution of binding activity, cross-competition of binding sites in EMSA, similar protein size as demonstrated by UV cross-linking, and sequence identity of one of the 3' alpha E-binding sites with a BSAP-binding site within the promoter of the sea urchin late histone gene H2A-2.2. These observations indicate that 3' alpha E is one of the mammalian targets for NF-HB (BSAP). Transient-transfection assays with chloramphenicol acetyltransferase gene constructs containing 3' alpha E and mutant 3' alpha E, in which one of the NF-HB binding sites was inactivated by site-specific mutagenesis, showed ca. five- to sixfold-enhanced activity of mutated 3' alpha E over parental 3' alpha E in B-cell lines (NF-HB+), while no significant difference was observed in plasmacytoma cells (NF-HB-). We conclude from these observations that NF-HB (BSAP) acts as a repressor of the mouse IgH 3' alpha E.
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PMID:NF-HB (BSAP) is a repressor of the murine immunoglobulin heavy-chain 3' alpha enhancer at early stages of B-cell differentiation. 849 73

It has been shown that the mouse histone H10 promoter contains a DNA element, composed of a direct repeat of the sequence GGTGACC separated by 7 nt, which is able to bind retinoic acid receptors and to modulate transcription of reporter genes following treatment with retinoic acid. We have now investigated whether this DNA motif is also responsive to thyroid hormone. We co-transfected CV-1 monkey kidney cells with chloramphenicol acetyltransferase (CAT) expression plasmids containing either 740 bp of the H10 wild-type promoter or five copies of the repeat element cloned in front of the thymidine kinase promoter and expression vectors for human thyroid hormone receptors (TRs) alpha or beta and retinoid X receptor alpha (RXR alpha). Treatment of transfected cells with triiodothyronine led to a dose-dependent increase in CAT activity. Transfection experiments with increasing amounts of expression vectors for either TR alpha or RXR alpha resulted in up to 6-fold enhancement of CAT transcription. Furthermore, point mutations within the half-sites of the response element of the H10 promoter, as well as deletions within the interspace region, lowered CAT activity to 60-80% of that of the wild-type control. Electrophoretic mobility shift assays showed that the repeat element was able to form retarded complexes with TR alpha homodimers, as well as with TR alpha-RXR alpha heterodimers. Our results suggest that thyroid hormone receptors are involved in the regulation of mouse histone H10 expression.
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PMID:Thyroid hormone receptors bind to the promoter of the mouse histone H10 gene and modulate its transcription. 855 62

In view of the wide chromosomal distribution of short alternating purine-pyrimidine sequences capable of adopting a number of superhelical stress-dependent structural configurations (left-handed helices and cruciforms), the question has been posed whether such sequences exert any functional effects in vivo. A series of eukaryotic expression vectors were constructed which contained C.G tracts of various lengths in the promoter region. It was shown that insertion of C.G tracts of 12-16 bp significantly increased the level of expression of the chloramphenicol acetyltransferase reporter gene. It was also demonstrated that the formation of additional activation complexes and the use of a preferred "face" or side of the DNA molecule is not responsible for the increased transcription which was observed upon insertion of the C.G tracts. Comparative assays of chromatin structure at the chimeric promoters indicate that the alternating C.G tracts adopt a structure which is incapable of binding histone proteins. These results strongly suggest that control of access to chromatin is involved in regulating the transcriptional activity of the chimeric promoters. Possible molecular bases for this phenomena are discussed.
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PMID:Enhancement of transcription by short alternating C.G tracts incorporated within a Rous sarcoma virus-based chimeric promoter: in vivo studies. 884 44

The sea urchin early histone repeating unit contains one copy of each of the five histone genes whose coordinate expression during development is regulated by gene-specific elements. To learn how within the histone repeating unit a gene-specific activator can be prevented to communicate with the heterologous promoters, we searched for domain boundaries by using the enhancer blocking assay. We focused on the region near the 3' end of the H2A gene where stage-specific nuclease cleavage sites appear upon silencing of the early histone genes. We demonstrated that a DNA fragment of 265 bp in length, defined as sns (for silencing nucleoprotein structure), blocked the enhancer activity of the H2A modulator in microinjected sea urchin embryos only when placed between the enhancer elements and the promoter. We also found that sns silenced the modulator elements even when placed at 2.7 kb from the promoter. By contrast, the enhancer activity of the modulator sequences, located downstream to the coding region, was not affected when sns was positioned in close proximity to the promoter. Finally, the H2A sns fragment placed between the simian virus 40 regulative region and the tk promoter repressed chloramphenicol acetyltransferase expression in transfected human cell lines. We conclude that 3' end of the H2A gene contains sequence elements that behave as functional barriers of enhancer function in the enhancer blocking assay. Furthermore, our results also indicate that the enhancer blocking function of sns lacks enhancer and species specificity and that it can act in transient assays.
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PMID:Enhancer blocking activity located near the 3' end of the sea urchin early H2A histone gene. 912 84

The testis-specific histone gene H1t is expressed only in mammalian testis at the stage of pachytene spermatocytes. The tissue-specific regulation of the mouse H1t gene was examined in mouse testicular primary culture cells with gene constructs consisting of H1t promoter elements fused to the chloramphenicol acetyltransferase or the firefly luciferase reporter gene. Our experiments demonstrate that expression of the mouse H1t gene is enhanced by a conserved H1 histone gene-specific TG box 452 base pairs upstream of the transcription start site. The transcription of the H1t gene appears to be reduced by sequences between -1999 and -1506. No regulatory effect could be shown for the H1 box in the expression of the mouse H1t gene. Binding of nuclear protein extracted from mouse testis to these consensus elements was shown by electrophoretic mobility-shift assays with mouse testicular nuclear proteins and labeled oligonucleotides containing the upstream TG box sequences or the two testis-specific elements of the H1t gene.
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PMID:Testis-specific expression of the mouse histone gene H1t is regulated by several promoter elements. 940 43

To compare the role of histone deactylation in estrogen activation of a transiently transfected vitellogenin (VIT) promoter and an integrated VIT promoter in the same cells, we produced three HepG2, human hepatoma, cell lines (HepG2ERV cells) stably expressing human estrogen receptor alpha (hERalpha) and containing an integrated VIT promoter-chloramphenicol acetyltransferase (VIT-CAT) reporter gene. The three ER-positive HepG2ERV cell lines and wild-type, ER-negative, HepG2 cells cotransfected with cytomegalovirus-hERalpha exhibited similar MOX-dependent inductions of 20- to 50-fold with a transiently transfected VIT-luciferase reporter and 15- to 50-fold with a transfected 4-estrogen response element-TATA-luciferase reporter gene. The histone deacetylase inhibitor, trichostatin A, did not enhance MOX induction of the transiently transfected VIT promoter in the HepG2ERV cells. In contrast, trichostatin A dramatically potentiated MOX induction of the stably integrated VIT-CAT reporter gene, resulting in MOX-ER-dependent increases in CAT activity of up to 600-fold. These data demonstrate that although liganded ER exhibits the capacity to fully activate a transiently transfected VIT promoter, under some circumstances the ability to reorganize a repressive chromatin structure may be limiting for steroid receptor action.
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PMID:A histone deacetylase inhibitor potentiates estrogen receptor activation of a stably integrated vitellogenin promoter in HepG2 cells. 1087 35

The induction of the beta interferon (IFN-beta) gene constitutes one of the first responses of the cell to virus infection. Its regulation is achieved through an intricate combination of virus-induced binding of transcription factors and local chromatin remodeling. In this work, we demonstrate that transcription factor YY1, known to interact with histone deacetylases (HDAC) and histone acetyltransferases, has a dual activator/repressor role during the regulation of the IFN-beta promoter activity. We show that YY1 specifically binds in vitro and in vivo to the murine IFN-beta promoter at positions -90 and -122. Overexpression of YY1 strongly repressed the transcriptional capacity of a stably integrated IFN-beta promoter fused to a chloramphenicol acetyltransferase reporter gene as well as the endogenous IFN activity of murine L929 cells via an HDAC activity. Stably integrated IFN-beta promoters mutated at the -90 site were no longer repressed by YY1, could no longer be activated by trichostatin A, displayed a retarded postinduction turn off, and a reduced virus-induced activity. Introduction of a mutation at the -122 site did not affect YY1-induced repression, but promoters with this mutation displayed a reduced virus-induced activity. Stably integrated full-length promoters (from position -330 to +20) mutated at both YY1-binding sites displayed extremely reduced promoter activities. We conclude that YY1 has a dual activator/repressor role on IFN-beta promoter activity depending on its binding site and time after infection.
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PMID:Transcription factor YY1 binds to the murine beta interferon promoter and regulates its transcriptional capacity with a dual activator/repressor role. 1258 14

It is well established that steroid receptor function requires interaction with coactivators. However, the mechanisms through which steroid receptors elicit precise assembly of coactivator complexes and the way the steroid activation signal is transduced remain elusive. Using a T47D cell line stably integrated with a mouse mammary tumor virus-chloramphenicol acetyltransferase (MMTV-CAT) reporter, we demonstrate that specific steroid receptors exhibit preferential recruitment of SRC-1 family coactivators, which determines the subsequent recruitment of specific downstream coregulator molecules. Upon ligand treatment, progesterone receptor (PR) interacted preferentially with SRC-1, which recruited CBP and significantly enhanced acetylation at K5 of histone H4. In contrast, activated glucocorticoid receptor (GR) preferentially associated with SRC-2 (TIF-2/GRIP-1), which subsequently recruited pCAF and led to specific modification of histone H3, suggesting that specific coactivators recruit distinct histone acetyltransferases to modulate the transcription of steroid-responsive genes. Loss-of-function experiments further support the predicted roles of SRC-1 and SRC-2 in, respectively, PR- and GR-mediated transcription on the MMTV promoter. This study indicates that differential recruitment of coactivators by nuclear receptors determines the assembly of coactivator complexes on target promoters to mediate specific transcription signals.
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PMID:Progesterone and glucocorticoid receptors recruit distinct coactivator complexes and promote distinct patterns of local chromatin modification. 1274 80


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