Characterization and Mapping of the Bovine FBP1 Gene

The steroid hormone 20-hydroxyecdysone plays a key role in the induction and modulation of morphogenetic events throughout Drosophila development. Previous studies have shown that a heterodimeric nuclear receptor composed of the EcR and USP proteins mediates the action of the hormone at the transcriptional through binding to palindromic ecdysteroid mediates the action of the hormone at the transcriptional level through binding to palindromic ecdysteroid response elements (EcREs) such as those present in the promoter of the hsp27 gene or the fat body-specific enhancer of the Fbp1 gene. We show that in addition to palindromic EcREs, the EcR/USP heterodimer can bind in vitro with various affinities to direct repetitions of the motif AGGTCA separated by 1 to 5 nucleotides (DR1 to DR5), which are known to be target sites for vertebrate nuclear receptors. At variance with the receptors, EcR/USP was also found to bind to a DR0 direct repeat with no intervening nucleotide. In cell transformation assays, direct repeats DR0 to DR5 alone can render the minimum viral tk or Drosophila Fbp1 promoter responsive to 20-hydroxyecdysone, as does the palindromic hsp27 EcRE. In a transgenic assay, however, neither the palindromic hsp27 element nor direct repeat DR3 alone can make the Fbp1 minimal promoter responsive to premetamorphic ecdysteroid peaks. In contrast, DR0 and DR3 elements, when substituted for the natural palindromic EcRE in the context of the Fbp1 enhancer, can drive a strong fat body-specific ecdysteroid response in transgenic animals. These results demonstrate that directly repeated EcR/USP binding sites are as effective as palindromic EcREs in vivo. They also provide evidence that additional flanking regulatory sequences are crucially required to potentiate the hormonal response mediated by both types of elements and specify its spatial and temporal pattern.

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In vivo functional characterization of an ecdysone response enhancer in the proximal upstream region of the Fbp1 gene of D. melanogaster

Mechanisms of Development ◽

10.1016/0925-4773(93)90062-3 ◽

1993 ◽

Vol 44 (2-3) ◽

pp. 123-138 ◽

Cited By ~ 14

Author(s):

Monique Laval ◽

Francine Pourrain ◽

Jean Deutsch ◽

Jean-Antoine Lepesant

Keyword(s):

Functional Characterization ◽

Upstream Region ◽

Fbp1 Gene

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The ecdysone response enhancer of the Fbp1 gene of Drosophila melanogaster is a direct target for the EcR/USP nuclear receptor.

Molecular and Cellular Biology ◽

10.1128/mcb.14.7.4465 ◽

1994 ◽

Vol 14 (7) ◽

pp. 4465-4474 ◽

Cited By ~ 47

Author(s):

C Antoniewski ◽

M Laval ◽

A Dahan ◽

J A Lepesant

Keyword(s):

Drosophila Melanogaster ◽

Binding Site ◽

Fat Body ◽

Sequence Similarity ◽

High Concentration ◽

Direct Role ◽

Palindromic Structure ◽

Fbp1 Gene

The transcription of the Drosophila melanogaster Fbp1 gene is induced by the steroid hormone 20-hydroxyecdysone and restricted to the late-third-instar fat body tissue. In a previous study we showed that the -68 to -138 region relative to the transcription start site acts as an ecdysone-dependent third-instar fat body-specific enhancer in a transgenic assay. Here we report that seven nucleoprotein complexes are formed in vitro on this enhancer when a nuclear extract from late-third-instar fat body is used in a gel shift assay. Accurate mapping of the binding sites of the complexes revealed a remarkably symmetrical organization. Using specific antibodies, one of the complexes was identified as a heterodimer consisting of the ecdysone receptor (EcR) and Ultraspiracle (USP) proteins. The binding site of the heterodimer as defined by mutagenesis and methylation interference experiments bears strong sequence similarity to the canonical hsp27 ecdysone response element, including an imperfect palindromic structure. The two elements diverge at three positions in both half-sites, indicating that the structure of an active EcR/USP binding site allows considerable sequence variations. In vivo footprinting experiments using ligation-mediated PCR and wild-type or ecdysteroid-deficient larvae show that occupancy of the Fbp1 EcR/USP binding site and adjacent region is dependent on a high concentration of ecdysteroids. These results provide strong evidence for a direct role of the EcR/USP heterodimer in driving gene expression in response to changes of the ecdysteroid titer during Drosophila larval development.

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Reciprocal stabilization of transcription factor binding integrates two signaling pathways to regulate fission yeast fbp1 transcription

Nucleic Acids Research ◽

10.1093/nar/gkab758 ◽

2021 ◽

Vol 49 (17) ◽

pp. 9809-9820

Author(s):

Wakana Koda ◽

Satoshi Senmatsu ◽

Takuya Abe ◽

Charles S Hoffman ◽

Kouji Hirota

Keyword(s):

Signal Transduction ◽

Transcription Factor ◽

Transcription Factors ◽

Fission Yeast ◽

Binding Sites ◽

Transcription Factor Binding ◽

Signal Transduction Pathways ◽

Factor Binding ◽

Close Proximity ◽

Fbp1 Gene

Abstract Transcriptional regulation, a pivotal biological process by which cells adapt to environmental fluctuations, is achieved by the binding of transcription factors to target sequences in a sequence-specific manner. However, how transcription factors recognize the correct target from amongst the numerous candidates in a genome has not been fully elucidated. We here show that, in the fission-yeast fbp1 gene, when transcription factors bind to target sequences in close proximity, their binding is reciprocally stabilized, thereby integrating distinct signal transduction pathways. The fbp1 gene is massively induced upon glucose starvation by the activation of two transcription factors, Atf1 and Rst2, mediated via distinct signal transduction pathways. Atf1 and Rst2 bind to the upstream-activating sequence 1 region, carrying two binding sites located 45 bp apart. Their binding is reciprocally stabilized due to the close proximity of the two target sites, which destabilizes the independent binding of Atf1 or Rst2. Tup11/12 (Tup-family co-repressors) suppress independent binding. These data demonstrate a previously unappreciated mechanism by which two transcription-factor binding sites, in close proximity, integrate two independent-signal pathways, thereby behaving as a hub for signal integration.

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Glucose repression of transcription of the Schizosaccharomyces pombe fbp1 gene occurs by a cAMP signaling pathway.

Genes & Development ◽

10.1101/gad.5.4.561 ◽

1991 ◽

Vol 5 (4) ◽

pp. 561-571 ◽

Cited By ~ 97

Author(s):

C S Hoffman ◽

F Winston

Keyword(s):

Schizosaccharomyces Pombe ◽

Signaling Pathway ◽

Glucose Repression ◽

Camp Signaling ◽

Camp Signaling Pathway ◽

Fbp1 Gene

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CCAAT-enhancer binding protein-α (C/EBPα) and hepatocyte nuclear factor 4α (HNF4α) regulate expression of the human fructose-1,6-bisphosphatase 1 (FBP1) gene in human hepatocellular carcinoma HepG2 cells

PLoS ONE ◽

10.1371/journal.pone.0194252 ◽

2018 ◽

Vol 13 (3) ◽

pp. e0194252 ◽

Cited By ~ 5

Author(s):

Siriluck Wattanavanitchakorn ◽

Pinnara Rojvirat ◽

Tanit Chavalit ◽

Michael J. MacDonald ◽

Sarawut Jitrapakdee

Keyword(s):

Hepatocellular Carcinoma ◽

Hepg2 Cells ◽

Binding Protein ◽

Human Hepatocellular Carcinoma ◽

Nuclear Factor ◽

Hepatocyte Nuclear Factor ◽

Enhancer Binding Protein ◽

Fructose 1,6 Bisphosphatase ◽

Hepatocyte Nuclear Factor 4Α ◽

Fbp1 Gene

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Deletion scanning of the regulatory sequences of the Fbp1 gene of Drosophila melanogaster using P transposase-induced deficiencies.

Genetics ◽

10.1093/genetics/135.3.801 ◽

1993 ◽

Vol 135 (3) ◽

pp. 801-816

Author(s):

P Lapie ◽

F Nasr ◽

J A Lepesant ◽

J Deutsch

Keyword(s):

Reporter Gene ◽

Fat Body ◽

Molecular Mapping ◽

Regulatory Element ◽

Regulatory Sequences ◽

Body Protein ◽

Cis Acting ◽

Genomic Location ◽

Adh Activity ◽

Fbp1 Gene

Abstract A procedure permitting deletion scanning of potential cis-regulatory sequences within a transgene whose genomic position remains fixed was applied to the study of the upstream sequences of the ecdysteroid-inducible Fat-body-protein-1 (Fbp1) gene. Deficiencies were induced in a Fbp1:Adh fusion transgene by means of a secondary P transposase mutagenesis. Phenotypic and molecular screens were used to select mutant transposons that retained their original genomic location and carried a deletion affecting the Fbp1 sequences but not the Adh reporter gene. Molecular mapping of the deletion breakpoints was achieved by sequence analysis and expression of the reporter gene was quantified by measurement of ADH activity. This procedure was efficient in detecting cis-acting elements, even those with moderate effects on levels of gene expression. For example, we have succeeded in identifying a negative regulatory element. Deletion of this element leads to a 50% increase in the reporter ADH activity. This element binds the transcription factor AEF-1. In addition, we have detected a strong, positively acting element contained within a 32-bp region located immediately upstream of an ecdysone-response element.

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A Natural A/T-Rich Sequence from the Yeast FBP1 Gene Exists as a Cruciform in Escherichia coli Cells

Plasmid ◽

10.1006/plas.1993.1024 ◽

1993 ◽

Vol 29 (3) ◽

pp. 222-232 ◽

Cited By ~ 4

Author(s):

Marcel.lı́ del Olmo ◽

José E. Pérez-Ortı́n

Keyword(s):

Escherichia Coli ◽

Escherichia Coli Cells ◽

Fbp1 Gene

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Fission Yeast Tup1-Like Repressors Repress Chromatin Remodeling at thefbp1+ Promoter and theade6-M26Recombination Hotspot

Genetics ◽

10.1093/genetics/165.2.505 ◽

2003 ◽

Vol 165 (2) ◽

pp. 505-515 ◽

Cited By ~ 1

Author(s):

Kouji Hirota ◽

Charles S Hoffman ◽

Takehiko Shibata ◽

Kunihiro Ohta§

Keyword(s):

Transcription Factor ◽

Fission Yeast ◽

Chromatin Remodeling ◽

Glucose Repression ◽

Regulation Of Gene Expression ◽

Regulatory Elements ◽

Chromatin Regulation ◽

Wild Type ◽

Responsive Element ◽

Fbp1 Gene

AbstractChromatin remodeling plays crucial roles in the regulation of gene expression and recombination. Transcription of the fission yeast fbp1+ gene and recombination at the meiotic recombination hotspot ade6-M26 (M26) are both regulated by cAMP responsive element (CRE)-like sequences and the CREB/ATF-type transcription factor Atf1•Pcr1. The Tup11 and Tup12 proteins, the fission yeast counterparts of the Saccharomyces cerevisiae Tup1 corepressor, are involved in glucose repression of the fbp1+ transcription. We have analyzed roles of the Tup1-like corepressors in chromatin regulation around the fbp1+ promoter and the M26 hotspot. We found that the chromatin structure around two regulatory elements for fbp1+ was remodeled under derepressed conditions in concert with the robust activation of fbp1+ transcription. Strains with tup11Δ tup12Δ double deletions grown in repressed conditions exhibited the chromatin state associated with wild-type cells grown in derepressed conditions. Interestingly, deletion of rst2+, encoding a transcription factor controlled by the cAMP-dependent kinase, alleviated the tup11Δ tup12Δ defects in chromatin regulation but not in transcription repression. The chromatin at the M26 site in mitotic cultures of a tup11Δ tup12Δ mutant resembled that of wild-type meiotic cells. These observations suggest that these fission yeast Tup1-like corepressors repress chromatin remodeling at CRE-related sequences and that Rst2 antagonizes this function.

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