Identification of promoter elements in 5'-flanking region of murine cyclic nucleotide phosphodiesterase 3B gene.

Item request has been placed! ×
Item request cannot be made. ×
loading   Processing Request
Read More Add to Saved list
  • Author(s): Liu H;Liu H; Tang JR; Degerman E; Manganiello VC
  • Source:
    Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2005; Vol. 307, pp. 109-24.
  • Publication Type:
    Journal Article
  • Language:
    English
  • Additional Information
    • Source:
      Publisher: Humana Press Country of Publication: United States NLM ID: 9214969 Publication Model: Print Cited Medium: Print ISSN: 1064-3745 (Print) Linking ISSN: 10643745 NLM ISO Abbreviation: Methods Mol Biol Subsets: MEDLINE
    • Publication Information:
      Publication: Totowa, NJ : Humana Press
      Original Publication: Clifton, N.J. : Humana Press,
    • Subject Terms:
      3',5'-Cyclic-AMP Phosphodiesterases/*genetics ; Response Elements/*genetics; 3',5'-Cyclic-AMP Phosphodiesterases/biosynthesis ; 3T3 Cells ; Activating Transcription Factor 2 ; Animals ; Cyclic AMP Response Element-Binding Protein/genetics ; Cyclic Nucleotide Phosphodiesterases, Type 3 ; Fibroblasts/metabolism ; Mice ; Response Elements/physiology ; TATA Box/genetics ; TATA Box/physiology ; Transcription Factors/genetics
    • Abstract:
      We describe techniques for identifying functional promoter elements in the 5'-flanking region of the murine cyclic nucleotide phosphodiesterase 3B (mPDE3B) gene. The 5'-flanking region of the mPDE3B gene was cloned and sequenced, and putative transcription factor binding sites were identified with computational tools. A series of reporter plasmids containing the luciferase gene fused to different fragments of the 5'-flanking region of the mPDE3B gene was constructed and used to transfect 3T3-L1 fibroblasts or differentiating adipocytes. Reporter gene assays showed that there are two promoter regions in the 5'-flanking region in the mPDE3B gene: a distal region located approx 4 kb upstream of the translation initiation site that contains cAMP-response element (CRE) cis-acting elements, and a proximal region that is GC rich and lacks TATA sequences. The distal promoter region induced much higher luciferase activity than did the proximal one. Mutation of the CRE sequences or reversal of the orientation of the CRE-containing region abolished promoter activity of the distal region. Electrophoretic mobility shift assay analysis indicated that binding to CRE elements was greater in nuclear extracts from differentiating adipocytes than from fibroblasts. Mapping of transcription initiation sites suggested that the distal promoter region might function as an enhancer, whereas the proximal promoter drives transcription of the mPDE3B gene.
    • Accession Number:
      0 (Activating Transcription Factor 2)
      0 (Cyclic AMP Response Element-Binding Protein)
      0 (Transcription Factors)
      EC 3.1.4.17 (3',5'-Cyclic-AMP Phosphodiesterases)
      EC 3.1.4.17 (Cyclic Nucleotide Phosphodiesterases, Type 3)
      EC 3.1.4.17 (Pde3b protein, mouse)
    • Publication Date:
      Date Created: 20050701 Date Completed: 20050915 Latest Revision: 20071115
    • Publication Date:
      20240829
    • Accession Number:
      10.1385/1-59259-839-0:109
    • Accession Number:
      15988059