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 5'-flanking region of human Cu/Zn superoxide dismutase (SOD1) was cloned from human genomic library for the study of regulation of human SOD1 gene. We determined 3678 nucleotide sequences of 5'-flanking region of human SOD1. The putative binding sites of transcriptional factors such as NF1, Sp1, AP1, AP2, GRE, HSE and NF kappa B were found. The upstream region of this gene was analyzed by deletion and measuring the linked chloramphenicol acetyltransferase (CAT) activities. Several deletion analyses of promoter activity indicated that there were positive and negative regulatory regions. The region from -1325 bp to -1040 bp was found to have a heat shock response element.
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PMID:Study of 5'-flanking region of human Cu/Zn superoxide dismutase. 802 98

The 230-kD bullous pemphigoid antigen is a hemidesmosomal protein of the cutaneous basement membrane zone. We have previously cloned overlapping cDNAs corresponding to the human 230-kD bullous pemphigoid antigen gene (BPAG1), located at the human chromosomal locus 6p11-12. Utilizing the cDNA clones, a genomic DNA lambda FIX II phage library was screened. Seven over-lapping genomic clones, spanning approximately 20 kb, were isolated. These clones were shown to contain the entire approximately 9-kb coding sequence of BPAG1, and it consisted of 22 separate exons which varied from 78 to 2,810 bp in size. Elucidation of 2.6 kb of 5'-flanking DNA was found to contain several putative transcriptional response elements, and development of promoter chloramphenicol acetyltransferase (CAT) reporter gene constructs allowed identification of putative cis-elements which confer keratinocyte-specific expression to the gene. In particular, a putative AP2-binding sequence (KRE2) in the position -(1,786-1778) was shown to be responsible for marked enhancement of the endogenous promoter, as well as of a heterologous thymidine kinase/CAT construct, activity in normal human keratinocytes. Normal human keratinocyte nuclear extracts contained a protein, designated as KTP1, which complexed with the KRE2 oligomer by gel mobility shift assays. UV cross-linking and Southwestern analysis suggested that KTP1 is a DNA-binding protein clearly distinct from AP2, and this protein may be responsible for the basal keratinocyte-specific expression of the BPAG1 gene.
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PMID:Molecular biology of the 230-kD bullous pemphigoid antigen. Cloning of the BPAG1 gene and its tissue-specific expression. 804 59

We have recently cloned and characterized the entire human 230-kDa bullous pemphigoid antigen gene, which is expressed at a relatively high level in the basal keratinocytes. A putative AP2 binding sequence (KRE2), identified in the position -1786 to -1778, was cloned in front of a heterologous thymidine kinase chloramphenicol acetyltransferase construct, and transient transfections of normal human keratinocytes indicated a marked enhancement of the promoter activity. Normal human keratinocyte nuclear extracts contained a protein, designated as keratinocyte transcriptional protein-1 (KTP-1), which complexed with the KRE2 oligomer when examined by gel mobility shift assays. This protein was not detected in human skin fibroblast or HeLa cell nuclear extracts that did, however, contain AP2. UV cross-linking studies and Southwestern analyses suggested that KTP-1 binds to DNA as a single polypeptide of approximately 110 kDa. These data suggest that KTP-1 is a DNA-binding protein clearly distinct from AP2, and this protein may be responsible for the basal keratinocyte-specific expression of the bullous pemphigoid antigen gene.
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PMID:Identification of a DNA-binding protein (keratinocyte transcriptional protein-1) recognizing a keratinocyte-specific regulatory element in the 230-kDa bullous pemphigoid antigen gene. 827 41

During endochondral bone formation, hypertrophic chondrocytes initiate synthesis of type X collagen. Previous studies have shown that regulation of this molecule is at the level of transcription. To further explore this regulation, we have studied a segment of the type X collagen gene extending from 562 base pairs (bp) upstream to 86 bp downstream of the transcriptional start site. We have studied this "proximal promoter region" by both structural analysis by DNase I in vivo footprinting and functional analysis by transient transfections. In type X collagen-expressing, hypertrophic chondrocytes, in vivo footprinting detected a fully protected TATA region flanked by hypersensitive sites but no other major protection. Type X collagen-negative cells (nonhypertrophic chondrocytes and tendon fibroblasts) showed major protection at a number of other sites, most notably an 8-bp region overlapping an AP2 site and a 9-bp region including the sequence CACACA. The importance of the proximal promoter region in restricting expression of type X collagen to hypertrophic chondrocytes was supported by transfection studies. A chloramphenicol acetyltransferase construct containing this region directed 5-10-fold higher chloramphenicol acetyltransferase expression in hypertrophic chondrocytes than in the other cell types. A 2.6-kilobase upstream fragment produced no additional effect. Thus, the proximal promoter region contains at least some regulatory elements for the cell-specific expression of type X collagen.
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PMID:Tissue-specific regulation of the type X collagen gene. Analyses by in vivo footprinting and transfection with a proximal promoter region. 853 1

Our previous study has shown that chicken c-ros is specifically expressed in certain epithelial cells of kidney, intestine, lung, bursa, thymus, and testis, and the expression is regulated temporally and spatially. To explore the molecular basis for the regulation of c-ros expression, we have cloned and characterized the chicken c-ros promoter. The most 5' c-ros cDNA was isolated and sequenced. Using the 5' cDNA as a probe, three genomic DNA clones containing the 5' c-ros cDNA sequence were isolated. Primer extension and RNase protection analysis were used to map the transcription initiation site for the c-ros mRNA in kidney and intestine. The sequence of the 1.3-kb region upstream of the initiation site contains TATA and CAAT boxes at 26 and 54 nucleotides, respectively, upstream of the initiation site. In addition, transcription factor binding sites for AP1, AP2, and Oct1 and several direct and inverted repeats are present within 1 kb upstream of the initiation site. The 1.3-kb DNA, when placed upstream of the chloramphenicol acetyltransferase gene, was shown to be functionally active. Serial deletions of this putative c-ros promoter allowed us to define a minimum c-ros promoter and to identify positive and negative regulatory regions. Using two oligonucleotides corresponding to a positive regulatory and potential factor binding region, we have demonstrated, by gel mobility shift experiments, their specific binding to nuclear extracts from kidney, intestine, and thymus. The binding pattern corresponds to the tissue specificity and temporal control of c-ros mRNA expression.
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PMID:Cloning and functional characterization of the chicken c-ros promoter. 901 57

ChAT (choline acetyltransferase) is the enzyme responsible for acetylcholine synthesis and is specifically expressed in cholinergic neurons. To further characterize the transcriptional regulation of the hCHAT (human ChAT) gene by NGF, we examined the effects upon ChAT promoter activity of a family of transcription factors which are activated by NGF and several extracellular stimuli and encoded by immediate-early genes. These include NGFI-A (Egr1, zif268), NGFI-C (Egr2), Krox-20 and NGFI-B (Nurr77). Two fragments of the hChAT gene were used for functional analysis carrying 944 bp (P1) and 4000 bp (P1 + P2) of the 5' flanking region in front of the chloramphenicol acetyltransferase (CAT) reporter gene. They were transiently co-transfected with NGFI-A, NGFI-C, Krox-20 and NGFI-B expression vectors in NG108-15, SN6 and COS-1 cells. CAT activity after transfection of the p4000 ChAT-CAT reporter into both neuronal cell lines (NG108-15 and SN6 cells) was increased up to 5-fold in the presence of co-transfected NGFI-A and up to 5- and 12-fold after co-transfection of NGFI-C expression vector in NG108-15 and SN6 cells, respectively. In NG108-15 cells, dbcAMP excerted a strong enhancing activity on the transactivation properties of NGFI-C while this was not observed when cells were transfected with NGFI-A. These trans-activation effects were specific for neuronal cells. When NG108-15 cells were treated with dbcAMP in the presence of H89, a specific PKA inhibitor, the increase of transcriptional activity of NGFI-C was abolished, indicating that a signalling transduction mechanism through PKA plays a role in NGFI-C-induced trans-activation. Electrophoretic mobility-shift assays showed that the sequence GCCCGGGGAG (NGFRE) located 1205 bp upstream of the first coding ATG (E1) can bind NGFI-A but not NGFI-C. Several possibilities explaining the observed results are discussed. Finally, transfections of ChAT-CAT reporters including the P1 + P2 region or a minimal ChAT enhancer present in the P2 region in front of a heterologous promoter indicated the presence of a regulatory element which conferred AP2-dependent trans-activation with homologous as well as with heterologous promoter constructs.
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PMID:Transcriptional activation of human choline acetyltransferase by AP2- and NGF-induced factors. 938 76

Seventeen kilobases of genomic DNA containing the promoter and the coding region of the round-spotted pufferfish JAK1 gene was isolated and completely sequenced. This gene consists of 25 exons and 24 introns spanning about 13.5 kb, compared to > 30kb in carp JAK1 gene. Primer extension analysis revealed one transcription initiation site which was 376 bp upstream of the translation initiation site. The sequence of the 2.9 kb region upstream of the transcription initiation site contains numerous potential binding sites for transcription factors including HNF-5, GCF, Sp1, CRE, AP2, GATA, GAGA, E2A, p53, and NF-IL6. When this region was placed upstream of the chloramphenicol acetyltransferase (CAT) reporter gene and transfected into a carp CF cell line, it could drive the synthesis of CAT enzyme three times more efficiently than could the common carp JAK1 promoter.
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PMID:Genomic organization and characterization of the promoter region of the round-spotted pufferfish (Tetraodon fluviatilis) JAK1 kinase gene. 943 51

Galactocerebrosidase (GALC) is the lysosomal enzyme deficient in human and certain animal species with globoid cell leukodystrophy (GLD) or Krabbe disease. It catalyzes the hydrolysis of specific galactolipids including galactosylceramide and psychosine. The GALC protein is found in very low amounts in all tissues, which delayed its purification and the subsequent cloning of its cDNA and gene. We previously published the exon-intron organization of the human gene, but did not functionally analyze the 5' flanking region. We now provide a description of this GC-rich region which includes one potential YY1 element and one potential SP1 binding site. There are 13 GGC trinucleotides within the first 150 bp preceding the initiation codon. The 5' end of intron 1 contains six potential Sp1 binding sites, one AP1 binding site, and eight AP2 binding sites. A construct containing nucleotides -176 to -24 had the strongest promoter activity using a vector containing the chloramphenicol acetyltransferase reporter gene. We also provide evidence for the presence of inhibitory sequences located immediately upstream of the promoter region, and within the first 234 nucleotides of intron 1. These elements together with a suboptimal nucleotide at position +4 may explain the low level of GALC protein in all cell types.
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PMID:Analysis of the 5' flanking region of the human galactocerebrosidase (GALC) gene. 944 67

Lactoferrin, a ferric binding glycoprotein found in milk, can possibly prevent microbial infection of the mammary gland and gastrointestinal tract. To define the regulation of the porcine lactoferrin gene (pLTF), we cloned its 5'-flanking region from a porcine liver genomic library and analyzed the 5' upstream region of approx. 4kb, two exons, and an intron. The transcription start site was localized by primer extension to residue G, which is 41 nucleotides upstream from the ATG start codon. The pLTF 5'-flanking region possesses several putative cis-acting regulatory elements found in both housekeeping and inducible genes; to define their function, they were inserted into a chloramphenicol acetyltransferase reporter construct. The region up to -156 sufficed for basic promoter activity, whereas the region up to -780 was required for maximal promoter activity in porcine testis cells (STcells), kidney cells (PK15 cells) and human mammary epithelial cells (HBL-100 cells). Detailed analysis of this proximal region by DNase I footprinting and electrophoretic mobility shift assays reveals that the ubiquitous factors SP1, AP2 and the mammary gland-specific factor (MGF) might play significant roles in regulating the transcription of the pLTF gene.
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PMID:Characterization and functional analysis of the porcine lactoferrin gene promoter. 966 28

A single gene, SIAT1, encodes ST6Gal I, the sialyltransferase that mediates transfer of alpha2,6-linked sialic acids to Galbeta1, 4GlcNAc termini of N-linked glycoproteins. In vivo, multiple SIAT1 mRNA forms, differing only in the 5'-untranslated region, are expressed in a tissue-specific manner. This mRNA heterogeneity has been attributed, at least in part, to transcription from a number of physically distinct promoter regions. In mature B-lymphocytes, SIAT1 transcription initiates at P2, a regulatory region known to function only in B-lineage cells. Bacterial chloramphenicol acetyltransferase (CAT) under the control of the P2 region encompassing 415 bp 5'- and 125 bp 3' of the transcriptional initiation site is efficiently expressed in Louckes, a mature B-lymphoblastoid cell line. In contrast, CAT expression in Reh, a T-null/B-null precursor line, and in HepG2, a hepatoma line, are 14-fold and >25-fold less than in Louckes, respectively. The data is consistent with the presence of cis -acting regulatory elements residing both 5' and 3' of the P2 transcriptional initiation site. At least 370 bp of 5'-flanking sequence, coinciding with the inclusion of AP2 and NF-kappaB sites, is necessary for high level expression in Louckes. Exon sequences 3' of the transcription start site are also important for expression. A segment from(+)32 to(+)125 (position(+)1 is transcription start site) is capable of exerting promoter-like activity in Louckes, but not in Reh or HepG2. CAT expression by P2 is negligible in Reh cells. However, enhanced CAT activity is not accompanied by elevated mRNA levels. This observation is consistent with the relief of translational restraints imposed by the(+)32 to(+)125 region. Together, the data demonstrate that efficient and cell-specific transcription regulation in mature B lymphocytes is contained in a 495 bp P2 segment that is comprised of 370 bp of 5'-flanking region and 125 bp of transcribed region of Exon X.
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PMID:Transcription of the beta-galactoside alpha2,6-sialyltransferase gene (SIAT1) in B-lymphocytes: cell type-specific expression correlates with presence of the divergent 5'-untranslated sequence. 1046 Aug 32


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