Transcriptional activation of the minimal human Proα1(I) collagen promoter: obligatory requirement for Sp1

A construct containing human Proα1(I) collagen gene promoter/enhancer-driven chloramphenicol acetyltransferase (CAT), pCOL-KT, failed to be expressed significantly in Sp1-deficient Schneider Drosophila line 2 (SL2) cells. However, CAT expression was induced 200-fold in SL2 cells co-transfected with pCOL-KT and pPACSp1, an Sp1-expression vector driven by the Drosophila actin 5C promoter. Elimination of the four potential Sp1-binding sites from pCOL-KT (pCOL-KTΔI), by removal of the first intron, did not abrogate Sp1-mediated induction of CAT. Even more significantly, a minimal Proα1(I) collagen promoter (-100 to +117 bp), containing a TATA box (-28 to -25 bp) and one putative Sp1-binding site (-87 to -82 bp), elicited strong Sp1-induced transactivation. Furthermore, mutation of the Sp1 motif in the minimal Proα1(I) collagen promoter-CAT construct abolished Sp1-induced expression of the reporter gene. Purified Sp1 protein bound specifically to DNA fragments of the Proα1(I) minimal promoter encompassing the putative Sp1-binding site; Sp1 binding could be competed out by a double-stranded oligonucleotide containing the wild-type Sp1 sequence, while an oligonucleotide containing a mutated Sp1 site failed to compete. Based on these results, we postulate that Sp1 plays an obligatory role in the transcriptional activation of the human Proα1(I) collagen gene. Additionally, we propose that a bona fide Sp1 motif, located most proximal to the TATA box, is necessary and sufficient for Sp1-mediated activation of the minimal Proα1(I) collagen promoter.

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Single-strand-DNA-binding factors specifically recognize the pyrimidine element in the chick α2(I) collagen gene promoter

Biochemical Journal ◽

10.1042/bj3140293 ◽

1996 ◽

Vol 314 (1) ◽

pp. 293-296 ◽

Cited By ~ 8

Author(s):

Dashzeveg BAYARSAIHAN ◽

Lewis N. LUKENS

Keyword(s):

Dna Binding ◽

Chick Embryo ◽

Gene Promoter ◽

Binding Activity ◽

Dna Conformation ◽

Collagen Gene ◽

Nuclear Extracts ◽

Binding Factor ◽

Dna Binding Factors ◽

Collagen Promoter

A pyrimidine element with mirror repeats centred at position -192 bp of the chick α2(I) collagen promoter interacts with sequence-specific DNA-binding factors. These factors bind to only the pyrimidine strand of this region and have no affinity for the complementary purine strand. Binding activity is also seen with the double-stranded form of this element, but with less affinity than to the single-stranded pyrimidine species. Southwestern blot analyses have shown that proteins of 80 and 134 kDa in chick embryo fibroblast nuclear extracts bind to the pyrimidine strand, whereas only a 134 kDa DNA-binding protein was found in chondrocyte nuclear extracts. The binding mechanism of these nuclear proteins with single-stranded DNA might be based on a non-B-DNA conformation of the pyrimidine element. The position of this binding site in the promoter region, its potential for adopting an unusual secondary structure and the presence of the 80 kDa binding factor in chick embryo fibroblasts, but not in chondrocytes, suggest a possible role for this factor in the expression of the α2(I) collagen gene.

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A Nucleosome-Free dG-dC-Rich Sequence Element Promotes Constitutive Transcription of the Essential Yeast RIO1 Gene

Biological Chemistry ◽

10.1515/bc.2003.143 ◽

2003 ◽

Vol 384 (9) ◽

pp. 1287-1292 ◽

Cited By ~ 6

Author(s):

M. Angermayr ◽

K. Schwerdtfeger ◽

W. Bandlow

Keyword(s):

Transcriptional Activation ◽

Transcription Initiation ◽

Essential Gene ◽

Core Promoter ◽

Tata Box ◽

Promoter Element ◽

Serine Kinase ◽

Sequence Element ◽

Dna Binding Factors ◽

Necessary And Sufficient

AbstractRIO1 is an essential gene that encodes a protein serine kinase and is transcribed constitutively at a very low level. Transcriptional activation of RIO1 dispenses with a canonical TATA box as well as with classical transactivators or specific DNA-binding factors. Instead, a dG-dC-rich sequence element, that is located 40 to 48 bp upstream the single site of mRNA initiation, is essential and presumably constitutes the basal promoter. In addition, we demonstrate here that this promoter element comprises a nucleosomefree gap which is centered at the dG-dC tract and flanked by two positioned nucleosomes. This element is both, necessary and sufficient, for basal transcription initiation at the RIO1 promoter and, thus, constitutes a novel type of core promoter element.

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A nuclear factor 1 binding site mediates the transcriptional activation of a type I collagen promoter by transforming growth factor-β

Cell ◽

10.1016/s0092-8674(88)80033-3 ◽

1988 ◽

Vol 52 (3) ◽

pp. 405-414 ◽

Cited By ~ 413

Author(s):

Pellegrino Rossi ◽

Gerard Karsenty ◽

Anita B. Roberts ◽

Nanette S. Roche ◽

Michael B. Sporn ◽

...

Keyword(s):

Growth Factor ◽

Binding Site ◽

Transcriptional Activation ◽

Type I Collagen ◽

Transforming Growth Factor ◽

Transforming Growth Factor Β ◽

Type I ◽

Nuclear Factor ◽

Nuclear Factor 1 ◽

Collagen Promoter

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Sequential Action of Ets-1 and Sp1 in the Activation of the Human β-1,4-Galactosyltransferase V Gene Involved in Abnormal Glycosylation Characteristic of Cancer Cells

Journal of Biological Chemistry ◽

10.1074/jbc.m611862200 ◽

2007 ◽

Vol 282 (38) ◽

pp. 27702-27712 ◽

Cited By ~ 28

Author(s):

Takeshi Sato ◽

Kiyoshi Furukawa

Keyword(s):

Gene Expression ◽

Binding Site ◽

Cancer Cells ◽

Transcriptional Activation ◽

Promoter Activity ◽

A549 Cells ◽

Gene Promoter ◽

Dominant Negative ◽

Mobility Shift ◽

V Gene

Malignant transformation is associated with increased gene expression of β-1,4-galactosyltransferase (β-1,4-GalT) V, which contributes to the biosynthesis of highly branched N-linked oligosaccharides characteristic of cancer cells. Our previous study showed that expression of the human β-1,4-GalT V gene is regulated by Sp1 (Sato, T., and Furukawa, K. (2004) J. Biol. Chem. 279, 39574–39583), and a subsequent study showed that the gene expression is also activated by Ets-1, a product of the oncogene (Sato, T., and Furukawa, K. (2005) Glycoconj. J. 22, 365). Herein we report the mechanism of β-1,4-GalT V gene activation by these transcription factors. The gene expression and promoter activity of β-1,4-GalT V increased when the ets-1 cDNA was transfected into A549 cells, which contain a small amount of Ets-1, but decreased dramatically when the dominant-negative ets-1 cDNA was transfected into HepG2 cells, which contain a large amount of Ets-1. Luciferase assays using deletion constructs of the β-1,4-GalT V gene promoter showed that promoter region –116 to +22 is critical for the transcriptional activation of the gene by Ets-1. Despite the presence of one Ets-1-binding site, which overlapped the Sp1-binding site, electrophoretic mobility shift assays showed that the region bound preferentially to Sp1 rather than to Ets-1. To solve this problem, we examined the transcriptional regulation of the human Sp1 gene by Ets-1 and found that the gene expression and promoter activity of Sp1 are regulated by Ets-1 in cancer cells. Functional analyses of two Ets-1-binding sites in the Sp1 gene promoter showed that only Ets-1-binding site –413 to –404 is involved in the activation of the gene by Ets-1. These results indicate that Ets-1 enhances expression of the β-1,4-GalT V gene through activation of the Sp1 gene in cancer cells.

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CpG methylation at the USF-binding site is important for the liver-specific transcription of the chipmunk HP-27 gene

Biochemical Journal ◽

10.1042/bj20051802 ◽

2006 ◽

Vol 395 (1) ◽

pp. 203-209 ◽

Cited By ~ 36

Author(s):

Gen Fujii ◽

Yuki Nakamura ◽

Daisuke Tsukamoto ◽

Michihiko Ito ◽

Tadayoshi Shiba ◽

...

Keyword(s):

Binding Site ◽

Transcriptional Activation ◽

Methylation Status ◽

Gene Promoter ◽

Cpg Methylation ◽

Flanking Sequence ◽

Upstream Stimulatory Factor ◽

Cpg Dinucleotides ◽

E Box ◽

Cpg Dinucleotide

The chipmunk hibernation-specific HP-27 gene is expressed specifically in the liver and has a CpG-poor promoter. To reveal how the liver-specific transcription of the HP-27 gene is regulated, we performed yeast one-hybrid screening of a chipmunk liver cDNA library. A 5′-flanking sequence of the HP-27 gene, extending from −170 to −140 and containing an E-box (5′-CACGTG-3′), is essential for the liver-specific transcription of HP-27. We used this sequence as bait and found that a ubiquitously expressed transcription factor, USF (upstream stimulatory factor), bound to the E-box. In COS-7 cells, USF activated transcription from the HP-27 gene promoter. We then used bisulphite genomic sequencing to analyse the methylation status of the four CpG dinucleotides that lie in the 5′-flanking sequence of the HP-27 gene up to −450, to investigate how the ubiquitously expressed USF activates transcription of the HP-27 gene only in the liver, while its transcription is repressed elsewhere. The only difference in methylation in the tissues tested was in the CpG dinucleotide in the USF-binding site, which was hypomethylated in the liver, but highly methylated in the kidney and heart. The specific methylation of the CpG dinucleotide at the USF-binding site impeded both the binding of USF and its transcriptional activation of the HP-27 gene. Chromatin immunoprecipitation using anti-USF antibodies revealed that USF bound to the HP-27 gene promoter in the liver, but not in the kidney or heart. Thus CpG methylation at the USF-binding site functions in establishing and maintaining tissue-specific transcription from the CpG-poor HP-27 gene promoter.

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The binding site for the liver-specific transcription factor Tf-LF1 and the TATA box of the human transferrin gene promoter are the only elements necessary to direct liver-specific transcriptionin vitro

Nucleic Acids Research ◽

10.1093/nar/18.19.5717 ◽

1990 ◽

Vol 18 (19) ◽

pp. 5717-5721 ◽

Cited By ~ 9

Author(s):

Daniel Mendelzon ◽

François Boissier ◽

Mario M. Zakin

Keyword(s):

Transcription Factor ◽

Binding Site ◽

Gene Promoter ◽

Tata Box ◽

Specific Transcription Factor ◽

Human Transferrin ◽

Transferrin Gene

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Transcriptional activation of USP16 gene expression by NFκB signaling

Molecular Brain ◽

10.1186/s13041-019-0535-3 ◽

2019 ◽

Vol 12 (1) ◽

Cited By ~ 2

Author(s):

Shou Yang ◽

Juelu Wang ◽

Shipeng Guo ◽

Daochao Huang ◽

Isabel Bestard Lorigados ◽

...

Keyword(s):

Gene Expression ◽

Down Syndrome ◽

Transcriptional Activation ◽

Mrna Level ◽

Gene Promoter ◽

Luciferase Assay ◽

Bona Fide ◽

Flanking Region ◽

Nfκb Pathway

AbstractUbiquitin Specific Peptidase 16 (USP16) has been reported to contribute to somatic stem-cell defects in Down syndrome. However, how this gene being regulated is largely unknown. To study the mechanism underlying USP16 gene expression, USP16 gene promoter was cloned and analyzed by luciferase assay. We identified that the 5′ flanking region (− 1856 bp ~ + 468 bp) of the human USP16 gene contained the functional promotor to control its transcription. Three bona fide NFκB binding sites were found in USP16 promoter. We showed that p65 overexpression enhanced endogenous USP16 mRNA level. Furthermore, LPS and TNFα, strong activators of the NFκB pathway, upregulated the USP16 transcription. Our data demonstrate that USP16 gene expression is tightly regulated at transcription level. NFκB signaling regulates the human USP16 gene expression through three cis-acting elements. The results provide novel insights into a potential role of dysregulation of USP16 expression in Alzheimer’s dementia in Down Syndrome.

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Repression of activator-mediated transcription by herpes simplex virus ICP4 via a mechanism involving interactions with the basal transcription factors TATA-binding protein and TFIIB.

Molecular and Cellular Biology ◽

10.1128/mcb.15.7.3618 ◽

1995 ◽

Vol 15 (7) ◽

pp. 3618-3626 ◽

Cited By ~ 33

Author(s):

B Gu ◽

R Kuddus ◽

N A DeLuca

Keyword(s):

Herpes Simplex Virus ◽

Herpes Simplex ◽

Dna Binding ◽

Binding Site ◽

Transcriptional Activation ◽

Transcription Initiation ◽

Binding Protein ◽

Tata Box ◽

Basal Transcription ◽

Simplex Virus

Infected-cell polypeptide 4 (ICP4) of herpes simplex virus is both a transcriptional activator and a repressor. It has been previously demonstrated that both SP1-activated transcription and USF-activated transcription are repressed by ICP4 without affecting basal transcription (B. Gu, R. Rivera-Gonzalez, C. A. Smith, and N. A. DeLuca, Proc. Natl. Acad. Sci. USA 90:9528-9532, 1993; R. Rivera-Gonzalez, A. N. Imbalzano, B. Gu, and N.A. DeLuca, Virology 202:550-564, 1994). In this study, it was found that ICP4 repressed the activation function of two other activators, VP16 and ICP4 itself, in vitro. ICP4 inhibited transcription by interfering with the formation of transcription initiation complexes without affecting transcription elongation. Repression of activator function required that an ICP4 DNA binding site was present in one orientation within approximately 45 bp 3' to the TATA box. DNA binding by ICP4 was necessary but not sufficient for repression. ICP4 has been shown to form tripartite complexes cooperatively with the TATA box-binding protein and TFIIB on DNA containing an ICP4 binding site and a TATA box (C. A. Smith, P. Bates, R. Rivera-Gonzalez, B. Gu, and N. DeLuca, J. Virol. 67:4676-4687, 1993). A region of ICP4 that enables the molecule to form tripartite complexes was also required in addition to the DNA binding domain for efficient repression. Moreover, repression was observed only when the ICP4 binding site was in a position that resulted in the formation of tripartite complexes. Together, the data suggest that ICP4 represses transcription by binding to DNA in a precise way so that it may interact with the basal transcription complex and inhibit some general step involved in the function of activators. The steps or interactions involved in transcriptional activation that are inhibited by ICP4 are discussed.

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