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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | De La Brousse, F. C. Shan, B. Chen, J. L. |
| Description | Author Affiliation: de la Brousse FC ( Tularik Inc., South San Francisco, CA 94080, USA.); |
| Abstract | Primer extension and RACE (rapid amplification of cDNA ends) assays were used to identify and sequence the 5' terminus of mouse ob mRNA. This sequence was used to obtain a recombinant bacteriophage containing the first exon of the encoding gene. DNA sequence analysis of the region immediately upstream of the first exon of the mouse ob gene revealed DNA sequences corresponding to presumptive cis-regulatory elements. A canonical TATA box was observed 30-34 base pairs upstream from the start site of transcription and a putative binding site for members of the C/EBP family of transcription factors was identified immediately upstream from the TATA box. Nuclear extracts prepared from primary adipocytes contained a DNA binding activity capable of avid and specific interaction with the putative C/EBP response element; antibodies to C/EBP alpha neutralized the DNA binding activity present in adipocyte nuclear extracts. When linked to a firefly luciferase reporter and transfected into primary adipocytes, the presumptive promoter of the mouse ob gene facilitated luciferase expression. When transfected into HepG2 cells, which lack C/EBP alpha, the mouse ob promoter was only weakly active. Supplementation of C/EBP alpha by cotransfection with a C/EBP alpha expression vector markedly stimulated luciferase expression. Finally, an ob promoter variant mutated at the C/EBP response element was inactive in both primary adipocytes and HepG2 cells. These observations provide evidence for identification of a functional promoter capable of directing expression of the mouse ob gene. |
| ISSN | 00278424 |
| e-ISSN | 10916490 |
| Journal | Proceedings of the National Academy of Sciences of the United States of America |
| Issue Number | 9 |
| Volume Number | 93 |
| Language | English |
| Publisher | National Academy of Sciences |
| Publisher Date | 1996-07-01 |
| Publisher Place | United States |
| Access Restriction | Open |
| Subject Keyword | Adipose Tissue Metabolism Obesity Genetics Promoter Regions, Genetic Proteins Animals Cell Nucleus Cloning, Molecular DNA Primers Exons Leptin Luciferases Biosynthesis Mice Mice, Inbred C57BL Mice, Obese Molecular Sequence Data Oligonucleotides, Antisense Polymerase Chain Reaction Protein Biosynthesis Recombinant Fusion Proteins Restriction Mapping TATA Box Transcription, Genetic Multidisciplinary |
| Content Type | Text |
| Resource Type | Article |
| Subject | Multidisciplinary |
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