Activation of gastrin gene transcription in islet cells by a RAP1‐like <i>cis</i>‐acting promoter element

Simon, Babette; Tillotson, Loyal G.; Brand, Stephen

doi:10.1016/0014-5793(94)00862-0

Cited by 6 publications

(15 citation statements)

References 39 publications

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“…(lanes 2-4; 1, 3, and 6 units/ml of the enzyme, respectively). After extraction, the DNA products of digestion were size-fractionated using gel electrophoresis, transferred to a nylon filter, and hybridized to the radioactively labeled probe C. consensus sequences as mammalian transcription factors (16,17). The recognition sequence of MIG1 (GCGGGG) is even present in multiple copies in the CpG island studied.…”

Section: Resultsmentioning

confidence: 99%

Nuclease-hypersensitive Chromatin Formed by a CpG Island in Human DNA Cloned as an Artificial Chromosome in Yeast

Mucha¹,

Lisowska²,

Goc³

et al. 2000

Journal of Biological Chemistry

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CpG islands are mostly unmethylated GC-, and CpGrich chromosomal segments overlapping promoter sequences in all housekeeping and many tissue-specific genes in vertebrates. Typically, these islands show an open chromatin structure, low in histone H1 and rich in acetylated histones. We have previously found that the island-like CGCG-rich sites in human DNA are hypersensitive to DNase I upon cloning in Saccharomyces cerevisiae. Here we studied, with a higher resolution, the chromatin formed in yeast by one such site, the CpG island accompanying the human glucose-6-phosphate dehydrogenase gene. We have found two strong hypersensitive sites and several positioned nucleosomes flanking the island despite the absence in yeast of such chromatin fiber-shaping factors as histone H1, methyltransferase, and the tissue-specific transcription factors. This finding, together with similar observations from our laboratories and others supports the idea that variations in GC and/or CpG content substantially contribute to the DNA sequence features modulating the structure of the chromatin. The composition-dependent fluctuations in the accessibility of DNA in the chromatin may constitute an evolutionary advantage and may explain the surprising compositional selection that acts in both the coding and non-coding segments of some genes during mammalian evolution.

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Section: Resultsmentioning

confidence: 99%

Nuclease-hypersensitive Chromatin Formed by a CpG Island in Human DNA Cloned as an Artificial Chromosome in Yeast

Mucha¹,

Lisowska²,

Goc³

et al. 2000

Journal of Biological Chemistry

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“…Both general and gene-speci®c transcription factors are highly conserved between yeast and mammalian cells (Buratowsky et al 1988;Chodosh et al 1988;Maity et al 1990;Simon et al 1994). Accordingly, we now report on a human gene which can functionally substitute for GCR2 in yeast.…”

Section: Introductionmentioning

confidence: 97%

A human gene, hSGT1, can substitute for GCR2, which encodes a general regulatory factor of glycolytic gene expression in Saccharomyces cerevisiae

Sato¹,

Jigami²,

Suzuki³

et al. 1999

Mol Gen Genet

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To determine whether similar regulatory mechanisms control the expression of glycolytic genes in yeast and human cells, we screened a human brain cDNA library for clones which complement the growth defect of the gcr2 mutant of Saccharomyces cerevisiae, and isolated hSGT1 (human suppressor of GCR two). Further work confirmed that the rescue of growth was associated with recovery of glycolytic enzyme activities, and that hSGT1 did not complement the growth defect of a gcr1 mutant. A hybrid protein comprising hSgt1p and the DNA-binding domain of Gal4p (GBD) activated a GAL1-lacZ reporter gene fusion, suggesting that the cloned gene may be a transcriptional activator. Two-hybrid experiments in yeast also indicate that hSgt1p interacts with Gcr1p. Northern analysis showed that hSGT1 is highly expressed in muscle and heart. Although the predicted amino acid sequence of hSgt1p does not display significant similarity to Gcr2p, we speculate that their functions may be analogous.

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“…Deletion constructs hugas119WT and hugas119mt were created using the polymerase chain reaction (PCR) and the hugas1300 construct as template. The hugas 119mt construct contains a single point mutation in the RAP1‐binding site as described [4]. Cohesive ends were generated on the PCR products by restriction digest before ligation into the pGL2‐basic vector.…”

Section: Methodsmentioning

confidence: 99%

“…Gene regulation is highly conserved between yeast and mammalian cells. We recently reported that the rat gastrin promoter contains a yeast RAP1‐binding site within the cis ‐regulatory domain GRD (gastrin regulatory domain) controlling gastrin gene transcription in islet cells [4]. Point mutations in the rat gastrin RAP1‐binding site abolished RAP1 binding and decreased transcriptional activation.…”

Section: Introductionmentioning

confidence: 99%

“…Recent data suggested that the CACC box in the human gastrin promoter, corresponding to the RAP1 site in the rat gastrin promoter, is also critical for gastrin activation in islet cells [5]. Furthermore, DNA–protein‐binding analyses revealed a DNA‐binding activity with RAP1‐like binding specificity in islet cells, suggesting a mammalian homologue RAP1‐like protein [4]. Gastrin presents an oncofetal expression pattern in islet cells.…”

Section: Introductionmentioning

confidence: 99%

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RAP1‐like binding activity in islet cells corresponds to members of the Sp1 family of transcription factors

et al. 1997

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Deletion and mutational analyses of the gastrin promoter have identified a binding site for the yeast transcription factor RAP1 relevant for transcriptional activation in islet cells. We here report that the mammalian transcription factors binding to this site in islet cells are the Sp transcription factor members Sp1 and Sp3. Furthermore, functional analyses revealed Sp1‐ and Sp3‐mediated transcriptional activation of gastrin. These data reveal that the zinc finger proteins Sp1 and Sp3 do have similar binding specificities as the multifunctional yeast RAP1 protein.

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Activation of gastrin gene transcription in islet cells by a RAP1‐like cis‐acting promoter element

Cited by 6 publications

References 39 publications

Nuclease-hypersensitive Chromatin Formed by a CpG Island in Human DNA Cloned as an Artificial Chromosome in Yeast

Nuclease-hypersensitive Chromatin Formed by a CpG Island in Human DNA Cloned as an Artificial Chromosome in Yeast

A human gene, hSGT1, can substitute for GCR2, which encodes a general regulatory factor of glycolytic gene expression in Saccharomyces cerevisiae

RAP1‐like binding activity in islet cells corresponds to members of the Sp1 family of transcription factors

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