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    Electronic Resource
    Electronic Resource
    Springer
    Fish physiology and biochemistry 20 (1999), S. 331-339 
    ISSN: 1573-5168
    Keywords: DNA response elements ; isomers ; nuclear receptors ; transcription regulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Transcription of growth hormone (GH) genes in vertebrates depends, among other factors, on triiodothyronine (T3) and retinoic acid (RA), mediated by nuclear receptors. In vitro studies of carp (Cyprinus carpio) pituitary cells have shown that addition of T3 and RA increase the steady state levels of the GH messenger RNA. In this study various isomers of fish thyroid hormone and retinoic acid receptor genes were tested in carp pituitary primary cell culture for transcription stimulation of Atlantic salmon (Salmo salar) GH promoter and of a general thyroid hormone-dependent promoter (4TREpTK). Zebrafish (Danio rerio) thyroid hormone receptor (TR) and retinoic acid receptor (RAR), in the presence of their ligands, T3 and RA respectively, increase synergistically expression of the reporter gene regulated by GH promoter, while only TR (and T3) is required for maximal stimulation from the 4TREpTK promoter. RA is essential for RAR mediated transcription from the GH promoter, while transcription stimulation by RAR from 4TREpTK promoter is ligand independent. Overexpression of the pituitary specific Japanese flounder (fl)TRβ isomer alone, or with either of the RAR isomers is more stimulatory than the TRα isomer in transcription from the pituitary specific endoTRE promoter element, while there is no preference among the TR or RAR isomers in stimulating transcription from the general consensus 4TREp. Furthermore, the recombinant TRα and TRα/RARα/γ heterodimers are shown to bind specifically to DNA elements. TRα binds specifically to endoTRE from Atlantic salmon GH promoter and to the consensus 4TREp. The zebrafish TRα and RAR(α/γ) preferentially heterodimerize on the consensus 4TREp, and the endoTRE successfully compete the complex formation. It would seem that the specific sequence of the promoter thyroid response elements controls the receptor isomer specificity, and the regulation of transcription by their ligands, thyroid hormone and RA.
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Fish physiology and biochemistry 17 (1997), S. 237-246 
    ISSN: 1573-5168
    Keywords: fish ; growth hormone gene ; thyroid hormone ; transcription ; DNA response element
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Transcription of growth hormone (GH) genes in vertebrates depends, among other factors, on pituitary specific transcription factors (such as Pit-1) and on thyroid hormone (TH) and TH receptors. Gel retardation experiments indicated the presence of proteins in the carp pituitary nuclear extract that specifically bind the Pit-1 DNA response element and the adjacent putative TH DNA response element (TRE), on the Atlantic salmon (Salmo salar) GH proximal promoter. The putative TRE element competes effectively in vitro in gel retardation assays with 4×TREpal, a consensus TRE element, on zebrafish (Danio rerio) purified recombinant TH receptor (zfTHR). The zfTHR/TRE complex can also be further identified with antibodies to rat THR that causes the complex a supershift in gel retardation assay. Cross competition was observed in nuclear extracts between the TRE and Pit-1 sites. It was found that 100 molar fold excess of Pit-1 binding element can compete effectively with the TH binding element and reduce its binding activity. The endogenous TRE binding element, on the other hand, cannot effectively reduce the binding activity of the Pit-1 DNA binding element. This unexpected competition between the two sites was examined in the presence and absence of the hormones: TH, 9-cis retinal, and TH + 9-cis retinal. The addition of the two hormones together significantly increased the binding properties of the TH response element as compared to its binding in the absence of the hormones, and also improved its binding activity in the presence of the Pit-1 DNA competitor. In summary, the two adjacent sites that control GH gene transcription, Pit-1 and TRE, on the fish GH promoter exhibit some degree of interaction between their respective transcription factors, the Pit-1 protein and the thyroid/retinoic acid receptors.
    Type of Medium: Electronic Resource
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