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Gene A4 is transcriptionally activated upon enterocyte differentiation of the human colonic epithelial cell line HT29-18 and its highly differentiated subclone HT29-18-C 1 [Oliva et al., Arch. Biochem. Biophys. 302 (1993) 183-192]. To characterize the mechanisms regulating the differential transcription of A4, we analyzed its immediate 5 -flanking region for regulatory elements. Promoter-linked...
Transcriptional regulation of the Chinese hamster ovary (CHO) adenine phosphoribosyltransferase-encoding gene (APRT) was studied. The 5 region of the CHO APRT is G + C-rich, but lacking TATA or CCAAT boxes. RNase protection assays indicate that it contains multiple transcription start points (tsp). A tsp 64 bp upstream from the translation start codon is denoted as +1. Linker-scanning (LS) mutation...
We report here the cloning, sequencing, functional analysis and DNase I footprinting of the human c-rel promoter region. The results revealed an 824-bp BsaAI-StuI minimal promoter region with a large number of NF-κB, Ap2 and Sp1-binding sites, some of them variants of known consensus sequences. This is the first promoter in the Rel/NF-κB/IκB family to be subjected to a detailed footprinting analysis...
We have identified a genomic clone containing the 5 regulatory region of the gene GTP-CH encoding human GTP cyclohydrolase I. The transcription start point (tsp) was mapped by 5 -rapid amplification of cDNA ends (5 -RACE). The 2.6-kb region upstream from the tsp showed promoter activity when ligated upstream from a reporter gene. The truncation of approximately 2 kb of the promoter did not...
Acyl-CoA-binding protein (ACBP) is an ubiquitously expressed 10-kDa protein which is present in high amounts in cells involved in solute transport or secretion. Rat ACBP is encoded by a gene containing the typical hallmarks of a housekeeping gene. Analysis of the promoter region of the rat ACBP gene by electrophoretic mobility shift assay (EMSA) revealed specific binding of proteins from rat liver...
Copper/zinc-containing superoxide dismutase (Cu/Zn SOD) is one of the essential enzymes that protects cells in aerobic conditions. To understand the mechanism of transcriptional regulation of the human Cu/Zn SOD gene, the proximal promoter region of Cu/Zn SOD has been investigated. In a previous study, it was found that transcription factors of the Sp1 and C/EBP family are associated with the expression...
Using rapid amplification of cDNA ends (RACE), we have cloned the 5 -untranslated region (5 -UTR) of the N-methyl-d-aspartate receptor subunit ε 2 from murine forebrain-derived mRNA. We identified two distinct types of cDNA species differing in the presence or absence of one exon sequence. Sequencing of the 5 -non-coding region of the ε 2 gene revealed that the ε 2 5 -UTR...
Filensin (also called CP94; CP95; CP97; 115kDa protein) is a component of the lens-specific beaded filament which is believed to be functionally important in lens fiber cell differentiation and in maintaining lens fiber cell conformation and transparency. A 17.2kb fragment containing the 5′-upstream sequence of the filensin gene was isolated. S1-mapping analysis determined the transcription start...
In previous work, we delineated a proximal region of the human α1(I) collagen gene (COL1A1) promoter necessary to direct its basal transcription in fibroblasts. This region has potential recognition sites for a variety of DNA-binding proteins. Here, we show that the -129/-107-bp sequence in this region of the promoter, which harbors an inverted CCAAT motif closely linked to a GC-rich direct repeat...
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...
Prosaposin is a multifunctional protein that encodes four glycoproteins, named saposins A, B, C and D. They participate in the catabolism of glycosphingolipids in lysosomes. When secreted, intact prosaposin may function as a neuritogenic factor. Human and mouse prosaposin displayed similar temporal and spatial regulation of expression. To gain insight into the transcriptional regulation of this locus,...
The mouse Ecm1 (extracellular matrix protein 1) gene codes for an extracellular protein of 85kDa. We have determined the primary structure of this gene and analysed 1665 bases of its 5' upstream regulatory region. The gene is approximately 5kb long and contains 11 exons. The exons range in size from 45 to 375bp, whereas the intron sizes ranges from 95 to 1115bp. All splice donor/acceptor sites conform...
The role of βAPP gene transcription and promoter regulation in modifying amyloid β-peptide (Aβ) levels is not well understood. Increased production of Aβ or changes in Aβ 42 /Aβ 40 ratio by fibroblasts occurs in the presence of mutant presenilin or βAPP alleles in familial Alzheimer's disease subjects. Both βAPP mRNA and Aβ levels are increased in trisomy 21. The APP gene...
Glutamic acid decarboxylase (GAD) 65 is one of two homologous proteins responsible for the synthesis of γ-aminobutyric acid, the most ubiquitous inhibitory neurotransmitter. In order to characterize the DNA elements responsible for controlling GAD65 expression, we cloned the 5' flanking region of the rat GAD65 gene. A major, proximal and a minor, distal region of transcription initiation were located...
GC-boxes and related motifs are frequently occurring DNA-elements present in many promoters and enhancers. In contrast to other elements it was generally thought that the transcription factor Sp1 is the only factor acting through these motifs. The cloning of paralogous genes of the Sp1 factor uncovered the existence of a small protein family consisting of Sp1, Sp2, Sp3 and Sp4. All four proteins exhibit...
Bcl-2-associated X protein (Bax) is a proapoptotic protein and is suggested to have an important role in carcinogenesis. To investigate the mechanism of bax gene transcriptional regulation, we isolated and sequenced the genomic DNA fragment of the 5' flanking region of the murine bax gene, and subcloned its promoter region into a luciferase reporter construction. The murine bax promoter is TATA-less,...
Embigin is a transmembrane glycoprotein belonging to the immunoglobulin superfamily, which is preferentially expressed in early stages of mouse embryogenesis and enhances integrin-mediated cell-substratum adhesion. The mouse embigin gene, which we cloned, spanned more than 50kb, in which nine exons were present. All exons contained protein-coding sequences. Each of the two immunoglobulin domains was...
Human prostate-specific transglutaminase (hTG P ) is a cross-linking enzyme encoded by the TGM4 gene. The TGM4 gene promoter was characterized by deletion mapping and mutational analysis. Promoter constructs, containing the minimal promoter requirements, could efficiently drive transcription in the prostate cancer cell lines PC346C and LNCaP and the hepatic cancer cell line Hep3B. The region...
Mutations in the gene encoding cystatin B (CSTB) are responsible for the primary defect in progressive myoclonus epilepsy of Unverricht-Lundborg type (EPM1). A novel and unique type of disease-causing mutation, an unstable dodecamer repeat expansion, accounts for the majority of EPM1 patients world-wide. This minisatellite repeat expansion, located in the putative promoter of CSTB 175 bp upstream...
Transcription factors of the basic Helix-Loop-Helix (bHLH) protein family play key roles in several developmental processes. Mist1 belongs to this group of proteins and shares several properties with the other family members. For example, Mist1 is capable of dimerization with the ubiquitously expressed E2A bHLH proteins and exhibits a strong DNA-binding activity to the core E-box sequence. Using...
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