References of "Muller, Marc"
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See detailAnalysis of the mouse doublecortin gene promoter in neurons
Piens, Marie; Muller, Marc ULg; Lion, Michelle ULg et al

Poster (2005)

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See detailEGF stimulates Pit-1 independent transcription of the human prolactin pituitary promoter in human breast cancer SK-BR-3 cells through its proximal AP-1 response element
Manfroid, Isabelle ULg; Van de Weerdt, Cécile ULg; Baudhuin, Ariane et al

in Molecular & Cellular Endocrinology (2005), 229(1-2), 127-39

Normal and neoplastic human mammary gland cells are targets for the proliferative action of prolactin. These cells also synthesize prolactin, thereby inducing an autocrine/paracrine proliferative loop. We ... [more ▼]

Normal and neoplastic human mammary gland cells are targets for the proliferative action of prolactin. These cells also synthesize prolactin, thereby inducing an autocrine/paracrine proliferative loop. We present the first extensive analysis of the transcriptional regulation of the human prolactin gene (hPRL) in human mammary tumor cells, SK-BR-3. We show that the pituitary promoter is functional in these cells in the absence of the pituitary-specific factor Pit-1. Expression of exogenous Pit-1 or epidermal growth factor (EGF) treatment stimulates the transfected hPRL pituitary promoter and the endogenous hPRL expression. EGF stimulation is mediated by increased synthesis of c-fos and c-jun, resulting in AP-1 binding to the proximal hPRL pituitary promoter. This regulation involves the EGF receptor, possibly ErbB2 that is highly expressed in SK-BR-3 cells, and a PI3K/JNK pathway. The stimulation of hPRL gene transcription by EGF in mammary cells may include hPRL in a complex regulatory network controlling growth of human mammary cells. [less ▲]

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See detailEnhancement of steroid receptor-mediated transcription for the development of highly responsive bioassays
Willemsen, Philippe; Scippo, Marie-Louise ULg; Maghuin-Rogister, Guy ULg et al

in Analytical and Bioanalytical Chemistry (2005), 382(4), 894-905

We have previously generated several transformed human mammary cell lines for the detection of steroid receptor-mediated activities and used these cell lines to detect and characterize steroid hormone ... [more ▼]

We have previously generated several transformed human mammary cell lines for the detection of steroid receptor-mediated activities and used these cell lines to detect and characterize steroid hormone (ant)agonistic compounds. In this report, we describe the specific optimization procedures used to enhance receptor-mediated transcription through the human glucocorticoid, progesterone and androgen receptors, respectively. Sodium arsenite-induced chemical stress leads to a substantial and specific increase in the glucocorticold receptor-mediated transcription, resulting in maximal stimulations of more than 2000-fold by the agonist dexamethasone. Similarly, a combined treatment with forskolin (an activator of adenylate cyclase) and trichostatin A (an inhibitor of histone deacetylases) leads to a synergistic enhancement of progesterone or androgen stimulation, resulting in a maximal induction of more than 200-fold or about 100-fold, respectively. The enhanced responses to specific steroids are mediated by the corresponding nuclear receptor. We show that by using these enhanced transcriptional stimulation protocols, it is possible to detect lower amounts of steroid hormones without substantially affecting the relative biological activities of various agonists. Finally, the application of these enhanced reporter cell assays to real biological samples from meat-producing animals is evaluated, and some validation parameters are presented. [less ▲]

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See detailUse of reporter cell lines for detection of endocrine-disrupter activity
Willemsen, Philippe; Scippo, Marie-Louise ULg; Kausel, G. et al

in Analytical and Bioanalytical Chemistry (2004), 378(3), 655-663

We have studied stable transformed human mammary cell lines with highly inducible steroid receptor-mediated luciferase reporter gene expression. Cells responding specifically to glucocorticoids ... [more ▼]

We have studied stable transformed human mammary cell lines with highly inducible steroid receptor-mediated luciferase reporter gene expression. Cells responding specifically to glucocorticoids, progestagens, androgens, or estrogens are described and characterized. The use of this high-throughput, cell-based assay for analysis of steroid (ant)agonists is reported. Systematic characterization of endocrine-disrupting activity on human receptors and in a human-cell system is interpreted for a selection of xenobiotics. We show that the phytoestrogens apigenin and genistin have progestagenic and androgenic activity, respectively. Finally, application of cell-based assays to the analysis of environmental samples is discussed. [less ▲]

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See detailRecombinant human estrogen, androgen and progesterone receptors for detection of potential endocrine disruptors
Scippo, Marie-Louise ULg; Argiris, Catherine; Van de Weerdt, Cécile ULg et al

in Analytical and Bioanalytical Chemistry (2004), 378(3), 664-669

This work reports the binding capacity of various chemicals (so-called endocrine disruptors) to recombinant human steroid receptors (hERalpha, hPR and hAR). The tested chemicals are organochlorine ... [more ▼]

This work reports the binding capacity of various chemicals (so-called endocrine disruptors) to recombinant human steroid receptors (hERalpha, hPR and hAR). The tested chemicals are organochlorine insecticides (DDT and its metabolites, methoxychlor, aldrin, dieldrin, chlordecone, lindane, trichlorobenzene), estrogenic insecticides (endosulfan, toxaphene, nonachlor), herbicides (alachlor and atrazine), fungicides (benomyl and vinclozolin), industrial chemicals (nonylphenol, bisphenol A, diphenylphtalate), antioxidants (butylated hydroxyanisol) and some phytoestrogens. Except for phytoestrogens, most of the tested chemicals (DDT and its metabolites, aldrin, alpha- and beta-endosulfan, toxaphen, trans-nonachlor) show higher affinities for hPR than for hERalpha, indicating that the interaction with the progesterone receptor could contribute to the endocrine-disrupting effects imputed to these chemicals. We propose to use binding assays using recombinant human steroid receptors as screening tools for the detection of endocrine disruptors in various samples. [less ▲]

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See detailReceptor-based screening assays: New perspectives in anti-doping control
Scippo, Marie-Louise ULg; Willemsen, Philippe; Danyi, Sophie ULg et al

in Chromatographia (2004), 59(Suppl. S), 23-27

The so-called "growth promoters", steroid hormones and beta-agonists, are currently controlled by using hyphenated analytical methods (chromatography coupled to mass spectrometry) or, sometimes for ... [more ▼]

The so-called "growth promoters", steroid hormones and beta-agonists, are currently controlled by using hyphenated analytical methods (chromatography coupled to mass spectrometry) or, sometimes for screening purposes, on immunoassays. These methods are often too specific to allow an effective multianalyte control. To develop more efficient assays, the use of hormonal receptors as detection tools (receptor-based binding assays and cell-based assays) is proposed. Receptor-based assays represent useful tools in screening of hormonal residues in food, but they could also be applied in doping control (to detect "new" hormonal substances). Furthermore, these assays could be used to monitor the human exposure to endocrine disruptors. [less ▲]

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See detailPitx factors are involved in basal and hormone-regulated activity of the human prolactin promoter
Quentien, M. H.; Manfroid, Isabelle ULg; Moncet, D. et al

in Journal of Biological Chemistry (2002), 277(46), 44408-44416

The pituitary-specific POU homeodomain factor Pit-1 likely interacts with other factors for cell-specific expression of prolactin. Here we identify the paired-like homeobox transcription factors Pitx1 and ... [more ▼]

The pituitary-specific POU homeodomain factor Pit-1 likely interacts with other factors for cell-specific expression of prolactin. Here we identify the paired-like homeobox transcription factors Pitx1 and Pitx2 as factors functionally activating the proximal human prolactin promoter (hPRL-164luc). Using in vitro binding assays and a series of site-specific mutations of the proximal hPRL promoter, we mapped the 131 and B2 bicoid sites involved in Pitx-mediated transactivation of the hPRL-164luc construct. In somatolactotroph GH4C1 cells, basal proximal hPRL promoter activity was inhibited by a Pitx2 dominant-negative form in a dose-dependent manner, whereas binding disruptive mutations in the Pitx sites significantly reduced basal activity of the promoter. We also show that synergistic activation of hPRL-164luc by Pitx2 and Pit-1 requires the integrity of the B2 Pitx binding site, and at least one of the P1 and P2 Pit-1 response elements. In addition, mutation in the B2 Pitx site results in attenuation of the promoter's responsiveness to forskolin, thyrotropin-releasing hormone, and epidermal growth factor. Conversely, Pitx1 or Pitx2 overexpression in GH4C1 cells leads to an enhancement of the drugs stimulatory effects. Altogether, these results suggest that full responsiveness to several signaling pathways regulating the hPRL promoter requires the B2 Pitx binding site and that Pitx factors may be part of the proteic complex involved in these regulations. Finally, in situ hybridization analysis showing coexpression of the PRL and Pitx2 genes in rat and human lactotroph cells corroborates the physiological relevance of these results. [less ▲]

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See detailTranscription of the human prolactin gene in mammary cells
Baudhuin, A.; Manfroid, Isabelle ULg; Van de Weerdt, Cécile ULg et al

in Annals of the New York Academy of Sciences (2002), 973

Expression of human prolactin in the Mammary gland, one of the main target organs of this hormone, leads to the formation of an autocrine-paracrine proliferative loop in this tissue. Involvement of ... [more ▼]

Expression of human prolactin in the Mammary gland, one of the main target organs of this hormone, leads to the formation of an autocrine-paracrine proliferative loop in this tissue. Involvement of prolactin in normal and neoplastic mammary development triggered the interest in transcriptional regulation of the human prolactin gene in mammary cells. Analysis of this regulation, and comparison to that in the pituitary, will contribute to a better understanding of mammary gland development and tumor formation. Here we present the first extensive analysis of the transcriptional regulation of the human prolactin gene in human mammary tumor cells. [less ▲]

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See detailA transformed fish cell line expressing a green fluorescent protein-luciferase fusion gene responding to cellular stress
Molina, A.; Carpeaux, Rudy ULg; Martial, Joseph ULg et al

in Toxicology in Vitro (2002), 16(2), 201-7

We obtained a stable transformed fish (EPC) cell line containing a reporter gene under the control of the tilapia HSP70 promoter. Expression of the reporter gene, coding for a green fluorescent protein ... [more ▼]

We obtained a stable transformed fish (EPC) cell line containing a reporter gene under the control of the tilapia HSP70 promoter. Expression of the reporter gene, coding for a green fluorescent protein (GFP)-luciferase fusion protein, was assessed by measuring the luciferase enzymatic activity by luminometry and the GFP expression by fluorescence microscopy and flow cytometry. The clone was characterized for its capacity to respond to heat shock treatment. The results show high induction after 1 h at 37 degrees C of treatment, up to 500-fold. In addition, its convenience to detect a large range of cellular stressors was evaluated. We observed high induction when Cd2+, Zn2+, Hg2+ or Cu2+ was added, but not Pb2+. In addition, activation of the reporter gene was observed in the presence of other compounds such as acetyl chloride, tetrachlorophenol, chloroacetamide and sodium arsenite. In conclusion, this cell line can be used as a rapid, cheap and easy biological test to determine cellular stress induced by environmental pollutants, alone or in conjunction with other, more specific assays. [less ▲]

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See detailExpression of the zinc finger Egr1 gene during zebrafish embryonic development
Close, Renaud; Toro, Sabrina; Martial, Joseph ULg et al

in Mechanisms of Development (2002), 118(1-2), 269-72

Egr1 is a highly conserved zinc finger protein which plays important roles in many aspects of vertebrate development and in the adult. The cDNA coding for zebrafish Egr1 was obtained and its expression ... [more ▼]

Egr1 is a highly conserved zinc finger protein which plays important roles in many aspects of vertebrate development and in the adult. The cDNA coding for zebrafish Egr1 was obtained and its expression pattern was examined during zebrafish embryogenesis using whole-mount in situ hybridization. Egr1 mRNA is first detected in adaxial cells in the presomitic mesoderm between 11 and 20 h post-fertilization (hpf), spanning the 4-24 somite stages. Later, Egr1 expression is observed only in specific brain areas, starting at 21 hpf and subsequently increasing in distinct domains of the central nervous system, e.g. in the telencephalon, diencephalon and hypothalamus. Between 24 and 48 hpf, Egr1 is expressed in specific domains of the hypothalamus, mesencephalon, tegmentum, pharynx, retina, otic vesicle and heart. [less ▲]

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See detailUse of specific bioluminescent cell lines for the detection of steroid hormone (ant)agonists in meat producing animals
Willemsen, Philippe; Scippo, Marie-Louise ULg; Maghuin-Rogister, Guy ULg et al

in Analytica Chimica Acta (2002), 473(1-2), 119-126

We present stable transformed human mammary cell lines displaying highly inducible steroid receptor-mediated luciferase reporter gene expression. The utilization of a cell-based assay for detection of ... [more ▼]

We present stable transformed human mammary cell lines displaying highly inducible steroid receptor-mediated luciferase reporter gene expression. The utilization of a cell-based assay for detection of steroid agonists and antagonists down to 0.3 ng g(-1) is described and its application to the analysis of bovine samples is discussed. In particular, the use of the assays to detect illegal progesterone or androgen treatment has to take into account the presence of natural endogenous hormones. (C) 2002 Elsevier Science B.V. All rights reserved. [less ▲]

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See detailDetection of illegal growth promoters in biological samples using receptor binding assays
Scippo, Marie-Louise ULg; Van de Weerdt, Cécile ULg; Willemsen, Philippe et al

in Analytica Chimica Acta (2002), 473(1-2), 135-141

In the European Union (EU), the use of growth-promoting substances in meat production is banned. The control of growth promoters, especially steroid hormones, is presently based on expensive and time ... [more ▼]

In the European Union (EU), the use of growth-promoting substances in meat production is banned. The control of growth promoters, especially steroid hormones, is presently based on expensive and time-consuming chromatographic methods of analysis or, sometimes, for screening purposes, on radio- or enzyme-immunoassays, all of which are often too specific to allow effective multi-analyte control. In order to develop rapid and inexpensive multi-analyte detection tests, we proposed the use of hormonal receptors as detection tools. The system described here (radio-receptor assays) is based on a direct bindin g assay of steroid hormones to their respective receptors. Human receptors to estrogens (hERalpha), androgens (hAR), progestagens (hPR) and glucocorticoids (hGR) have been produced by genetic engineering in bacteria or in eucaryotic cells. Binding analyses revealed that the obtained receptor proteins retained a high affinity for their corresponding native ligand. In addition, competition studies continued that each of the four receptors displays a specificity profile for a series of analogs in agreement with the literature. Finally, the stability of these recombinant receptors is sufficient to allow their use in test kits. (C) 2002 Elsevier Science B.V. All rights reserved. [less ▲]

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See detailHeat shock stimulation of a tilapia heat shock protein 70 promoter is mediated by a distal element
Molina, Alfredo; Di Martino, Emmanuel; Martial, Joseph ULg et al

in Biochemical Journal (2001), 356(Pt 2), 353-9

We reported previously that a tilapia (Oreochromis mossambicus) heat shock protein 70 (HSP70) promoter is able to confer heat shock response on a reporter gene after transient expression both in cell ... [more ▼]

We reported previously that a tilapia (Oreochromis mossambicus) heat shock protein 70 (HSP70) promoter is able to confer heat shock response on a reporter gene after transient expression both in cell culture and in microinjected zebrafish embryos. Here we present the first functional analysis of a fish HSP70 promoter, the tiHSP70 promoter. Using transient expression experiments in carp EPC (epithelioma papulosum cyprini) cells and in microinjected zebrafish embryos, we show that a distal heat shock response element (HSE1) at approx. -800 is predominantly responsible for the heat shock response of the tiHSP70 promoter. This element specifically binds an inducible transcription factor, most probably heat shock factor, and a constitutive factor. The constitutive complex is not observed with the non-functional, proximal HSE3 sequence, suggesting that both factors are required for the heat shock response mediated by HSE1. [less ▲]

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See detailGene structure and promoter function of a teleost ribosomal protein: a tilapia (Oreochromis mossambicus) L18 gene
Molina, Alfredo; Iyengar, Arati; Marins, Luis F. et al

in Biochimica et Biophysica Acta (2001), 1520(3), 195-202

We have cloned and characterized a tilapia (Oreochromis mossambicus) L18 ribosomal protein gene, including the complete transcribed region and 488 bp of upstream regulatory sequences. We have also ... [more ▼]

We have cloned and characterized a tilapia (Oreochromis mossambicus) L18 ribosomal protein gene, including the complete transcribed region and 488 bp of upstream regulatory sequences. We have also isolated two L18 cDNAs from another tilapia (Oreochromis niloticus) with a few conservative nucleotide differences. Our results suggest the presence of two genes in both species. Reporter constructs were tested for transient expression in CV1 cells and in microinjected zebrafish and tilapia embryos. The tilapia L18 promoter was able to drive expression of the reporter gene in all three experiments, with no apparent preference for a particular tissue. The tilapia L18 promoter is therefore likely to be a powerful tool to drive tissue-independent gene expression in fish. [less ▲]

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See detailThe ornithine decarboxylase gene is essential for cell survival during early murine development
Pendeville-Samain, Hélène ULg; Carpino, Nick; Marine, Jean-Christophe et al

in Molecular & Cellular Biology (2001), 21(19), 6549-58

Overexpression and inhibitor studies have suggested that the c-Myc target gene for ornithine decarboxylase (ODC), the enzyme which converts ornithine to putrescine, plays an important role in diverse ... [more ▼]

Overexpression and inhibitor studies have suggested that the c-Myc target gene for ornithine decarboxylase (ODC), the enzyme which converts ornithine to putrescine, plays an important role in diverse biological processes, including cell growth, differentiation, transformation, and apoptosis. To explore the physiological function of ODC in mammalian development, we generated mice harboring a disrupted ODC gene. ODC-heterozygous mice were viable, normal, and fertile. Although zygotic ODC is expressed throughout the embryo prior to implantation, loss of ODC did not block normal development to the blastocyst stage. Embryonic day E3.5 ODC-deficient embryos were capable of uterine implantation and induced maternal decidualization yet failed to develop substantially thereafter. Surprisingly, analysis of ODC-deficient blastocysts suggests that loss of ODC does not affect cell growth per se but rather is required for survival of the pluripotent cells of the inner cell mass. Therefore, ODC plays an essential role in murine development, and proper homeostasis of polyamine pools appears to be required for cell survival prior to gastrulation. [less ▲]

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See detailInhibition of protein phosphatase PP1 in GH3B6, but not in GH3 cells, activates the MEK/ERK/c-fos pathway and the human prolactin promoter, involving the coactivator CPB/p300
Manfroid, Isabelle ULg; Martial, Joseph ULg; Muller, Marc ULg

in Molecular Endocrinology (Baltimore, Md.) (2001), 15(4), 625-37

The human (hPRL) PRL gene proximal promoter (-164/+15) is the target for numerous signal transduction pathways involving protein kinases. The inhibitor of Ser/Thr-protein phosphatases okadaic acid (OA ... [more ▼]

The human (hPRL) PRL gene proximal promoter (-164/+15) is the target for numerous signal transduction pathways involving protein kinases. The inhibitor of Ser/Thr-protein phosphatases okadaic acid (OA) was shown to induce this promoter in rat pituitary GH3B6 through a synergism between increased amounts of the ubiquitous factor AP-1 and the pituitary-specific factor Pit-1. Here we show that this activation results mainly from transcriptional stimulation of the c-fos promoter leading to increased AP-1 activity. We report the surprising absence of the hPRL and c-fos promoter stimulation by OA in GH3 cells, closely related to GH3B6 cells, and we use this discrepancy to dissect the precise mechanism of action. c-fos gene activation involves the mitogen-activated kinase (MAPK)-ternary complex factor (TCF) pathway and can be obtained by expressing active V12ras in both cell lines. We show that OA acts by inhibiting protein phosphatase PP1, thereby protecting MAPK kinase (MEK)1/2 and/or a MEK1/2-kinase from dephosphorylation. PP1 inhibition of MEK activation by V12ras does not occur in GH3 cells, indicating that a distinct, PP1-sensitive phosphorylation site is used in GH3B6 cells to activate the TCF pathway in GH3B6 cells. Finally, we show that the synergistic OA activation of the hPRL promoter by Pit-1 and AP-1 is independent of the Pit-1 transactivation domain and is mediated by the general coactivator (CRE-binding protein)-binding protein (CBP)/p300. [less ▲]

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See detailCloning and expression analysis of an inducible HSP70 gene from tilapia fish
Molina, Alfredo; Biemar, Frédéric; Muller, Ferenc et al

in FEBS Letters (2000), 474(1), 5-10

We isolated and characterized the tilapia (Oreochromis mossambicus) HSP70 gene, highly homologous to other HSP70 genes. A dramatic increase of tilapia HSP70 mRNA levels was observed after heat shock of ... [more ▼]

We isolated and characterized the tilapia (Oreochromis mossambicus) HSP70 gene, highly homologous to other HSP70 genes. A dramatic increase of tilapia HSP70 mRNA levels was observed after heat shock of whole animals in all organs tested. Reporter constructs were tested for transient expression in carp cells and in microinjected zebrafish embryos. The entire isolated regulatory region (-851/+157) was able to mediate heat shock inducible expression of the reporter gene, with no preference for a particular tissue. Our studies represent the first transcriptional analysis of a HSP70 promoter from fish, revealing a powerful tool to direct controlled, tissue-independent gene expression in fish. [less ▲]

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See detailSilencing subdomains of v-ErbA interact cooperatively with corepressors: involvement of helices 5/6
Busch, Kerstin; Martin, Bernd; Baniahmad, Aria et al

in Molecular Endocrinology (Baltimore, Md.) (2000), 14(2), 201-11

Members of the thyroid hormone receptor (TR) family act on vertebrate development and homeostasis by activating or repressing transcription of specific target genes in a ligand-dependent way. Repression ... [more ▼]

Members of the thyroid hormone receptor (TR) family act on vertebrate development and homeostasis by activating or repressing transcription of specific target genes in a ligand-dependent way. Repression by TR in the absence of ligand is mediated by an active silencing mechanism. The oncogene v-ErbA is a variant form of TR unable to bind hormone and thus acts as a constitutive repressor. Functional studies and mutation analysis revealed that the TR/v-ErbA silencing domain is composed of three silencing subdomains (SSD1-3) which, although nonfunctional individually, synergize such that silencing activity is restored when they are combined in a heteromeric complex. Here we demonstrate, using protein interaction assays in vitro and in vivo, that the inactive v-ErbA point mutant L489R within helix 5/6 in SSD2 fails to interact with the two corepressors N-CoR (nuclear receptor corepressor) or SMRT (silencing mediator of retinoic acid and thyroid hormone receptor). Furthermore, mutants in SSD1 and SSD3 exhibit a reduced corepressor recruitment corresponding to their weak residual silencing activity. In mammalian two-hybrid assays, only the combination of all three silencing subdomains, SSD1-3, leads to a cooperative binding to the corepressors N-CoR or SMRT comparable to that of the full-length v-ErbA repression domain. In conclusion, full silencing activity requires corepressor interaction with all three silencing subdomains, SSD1-3. Among these, SSD2 is a new target for N-CoR and SMRT and is essential for corepressor binding and function. [less ▲]

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See detailFar upstream sequences regulate the human prolactin promoter transcription
Van de Weerdt, Cécile ULg; Peers, Bernard ULg; Belayew, A. et al

in Neuroendocrinology (2000), 71(2), 124-37

The human prolactin gene is mainly expressed in pituitary lactotrope cells, but transcription from an alternative, far upstream promoter was detected in lymphoid, placental and mammary cells. We describe ... [more ▼]

The human prolactin gene is mainly expressed in pituitary lactotrope cells, but transcription from an alternative, far upstream promoter was detected in lymphoid, placental and mammary cells. We describe the transcriptional activity in rat pituitary cells of the complete region separating the two promoters, using transient transfection experiments. A far upstream activating region was only functional in combination with the prolactin promoter. DNaseI protection experiments revealed, in addition to binding sites for the pituitary-specific factor Pit-1, sites (e.g. SD1) for several ubiquitous factors and one lymphoid-specific factor (SD4). A single copy of the ubiquitous site SD1 or the lymphoid-specific site SD4 was unable to activate transcription of a heterologous promoter in pituitary cells. However, SD1 activated transcription in nonpituitary cells and SD4 was functional specifically in lymphoid cells. Five copies of a distal site (D8) activated transcription in each cell type tested. Gel retardation experiments show that this site binds the specific factor C/EBP in liver and a distinct factor in other cell types. Our results suggest that different elements within this large region direct specific expression from each promoter via a complex interplay between cell-specific and ubiquitous transcription factors. [less ▲]

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