DNA recognition element required for PUF-I mediated cell-type-specific transcription of the rat prolactin gene

Sharp, Zelton Dave; Helsel, Sharon; Cao, Zhaodan; Barron, E. A.; Sánchez, Yolanda

doi:10.1093/nar/17.7.2705

Cited by 18 publications

(10 citation statements)

References 17 publications

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“…Thirty mutated promoters which had base substitutions introduced into 16 of the 18 targeted positions were assayed by in vitro transcription using GH3 and nonpituitary nuclear extracts. The results showed that the sequence 5'-ATAnCA-3' located from -52 to -46 was critically important for GH3-specific prolactin gene transcriptional efficiency (24). In agreement with these functional studies, mutations within the ATATTCA sequence which decrease transcriptional efficiency also decrease PUF-I binding (24).…”

Section: Cis-acting Regulatory Elements Of the Rat Prolacfin Genesupporting

confidence: 66%

“…The results showed that the sequence 5'-ATAnCA-3' located from -52 to -46 was critically important for GH3-specific prolactin gene transcriptional efficiency (24). In agreement with these functional studies, mutations within the ATATTCA sequence which decrease transcriptional efficiency also decrease PUF-I binding (24). These functional and binding data identified the ATATTCA sequence as the PUF-I recognition element and linked the PUF-I/Pit-1 binding activity with transcriptional regulation.…”

Section: Cis-acting Regulatory Elements Of the Rat Prolacfin Genesupporting

confidence: 65%

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Regulation of cell‐type‐specific transcription and differentiation of the pituitary

Sharp

Cao

1990

BioEssays

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The transcription of rat prolactin and growth hormone genes in vitro requires a pituitary transcription factor, specific to certain cell types in the pituitary, which currently appears to be the PUF-I/Pit-1/GHF-1 protein. This factor binds to cis-regulatory elements in the 5' region of both genes and exerts a positive influence on transcription initiation presumably by interacting with general transcription factors. The PUF-I/Pit-1/GHF-1 transcriptional regulatory protein probably has an important role in not only the differentiation of the pituitary lactotroph/somatotroph cell lineage; it is also expressed in the early development of the nervous system but its function there is less well documented. It appears to be one member of a family of trans-activator proteins involved in differential gene expression in several cell types.

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Section: Cis-acting Regulatory Elements Of the Rat Prolacfin Genesupporting

confidence: 66%

Section: Cis-acting Regulatory Elements Of the Rat Prolacfin Genesupporting

confidence: 65%

Regulation of cell‐type‐specific transcription and differentiation of the pituitary

Sharp

Cao

1990

BioEssays

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“…A recent base substitution mutational study has strengthened this view (42). Among the eight Pit-1 binding sites observed in the human PRL distal region, only sites D2 and D6 contain the exact consensus sequence, while the others contain related sequences with one or even two mismatches (Table 1).…”

Section: Resultsmentioning

confidence: 97%

“…Several in vivo and in vitro studies performed on the rat PRL gene have indicated that 400 base pairs (bp) of its 5'-flanking sequence are sufficient to confer pituitary-specific expression (6,16,36,40). This is achieved through the binding to this region of a pituitary-specific factor named LSF-1 (16,20), Puf-1 (6,7,42), or Pit-i (27,36). For simplicity's sake, this factor will be referred to as Pit-i.…”

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confidence: 99%

Regulatory elements controlling pituitary-specific expression of the human prolactin gene.

Peers¹,

Voz²,

Monget³

et al. 1990

Mol. Cell. Biol.

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We have performed transfection and DNase I footprinting experiments to investigate pituitary-specific expression of the human prolactin (hPRL) gene. When fused to the chloramphenicol acetyltransferase (CAT) reporter gene, 5,000 base pairs of the 5'-flanking sequences of the hPRL gene were able to drive high cat gene expression in prolactin-expressing GH3B6 cells specifically. Deletion analysis indicated that this pituitaryspecific expression was controlled by three main positive regulatory regions. The first was located just upstream from the TATA box between coordinates -40 and -250 (proximal region). We have previously shown that three motifs of this region bind the pituitary-specific Pit-l factor. The second positive region was located in the vicinity of coordinates -1300 to -1750 (distal region). DNase I footprinting assays revealed that eight DNA motifs of this distal region bound protein Pit-l and that two other motifs were recognized by ubiquitous factors, one of which seems to belong to the AP-1 (jun) family. The third positive region was located further upstream, between -3500 and -5000 (superdistal region). This region appears to enhance transcription only in the presence of the distal region.

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“…By using DNase I protection analysis, three pituitary-specific factor binding sites, labelled footprint I (FPI), FPIII, and FPIV ( Fig. 1), have been localized to the proximal 210 bp of the proximal rat prolactin (rPRL) promoter (21,23,40,47,57,58). All three elements bind Pit-1/GHF-1, which is a homeodomain-containing protein that trans-activates both GH and PRL promoters and, more importantly, appears to be critical for pituitary stem cell commitment to the somatotroph, lactotroph, and thyrotroph cell lineages (13,38,41,59,63).…”

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confidence: 99%

Interaction of basal positive and negative transcription elements controls repression of the proximal rat prolactin promoter in nonpituitary cells.

Jackson¹,

Keech²,

Williamson³

et al. 1992

Mol. Cell. Biol.

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The proximal rat prolactin (rPRL) promoter contains three cell-specific elements, designated footprints I, III, and IV, which restrict rPRL gene expression to anterior pituitary lactotroph cells. Footprint II (-130 to -120) binds a factor, which we have termed F2F, present in pituitary and nonpituitary cell types. Here we demonstrate that a key role of the footprint II site is to inhibit rPRL promoter activity in nonpituitary cells, specifically, by interfering with the basal activating function of a vicinal element. Gene transfer analysis revealed 20-fold activation of the rPRL promoter in nonpituitary cell types when footprint II was either deleted or specifically mutated. Similar activation of the intact rPRL promoter was obtained by in vivo F2F titration studies. In GH4 rat pituitary cells, the footprint II inhibitory activity was masked by the redundant, positively acting cell-specific elements and was inhibitory only if the two upstream sites, footprints III and IV, were deleted. Deletion of the -112 to -80 region in the footprint II site-specific mutant background resulted in complete loss of rPRL promoter activity in both pituitary and nonpituitary cell types, mapping a basal activating element that is operative irrespective of cell type to this region. While the basal activating element imparted an activating function in a heterologous promoter assay, the footprint II sequence did not display any inherent repressor function and actually induced several minimal heterologous promoters. However, the inhibitory activity of the footprint II site was detected only if it was in context with the basal activating element.These data underscore the importance of ubiquitous activating and inhibitory factors in establishing cell-specific gene expression and further emphasize the complexity of the molecular mechanisms which restrict gene expression to specific cell types. We provide a novel paradigm to study rPRL promoter function and hormone responsiveness independently of lactotroph cell-specific requirements.Eukaryotic cells have evolved a striking capacity to completely alter their program of gene expression in response to temporal, developmental, and metabolic signals. Indeed, differential activation and simultaneous repression of specific genes appears to be critical for orderly development of highly specialized tissues (19,22,37,65). While the role of nuclear factors which activate novel patterns of tissuespecific gene transcription during development have been well documented (6,13,17,22,24,25,33,38,52,59,60)

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DNA recognition element required for PUF-I mediated cell-type-specific transcription of the rat prolactin gene

Cited by 18 publications

References 17 publications

Regulation of cell‐type‐specific transcription and differentiation of the pituitary

Regulation of cell‐type‐specific transcription and differentiation of the pituitary

Regulatory elements controlling pituitary-specific expression of the human prolactin gene.

Interaction of basal positive and negative transcription elements controls repression of the proximal rat prolactin promoter in nonpituitary cells.

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