The mouse calretinin gene promoter region: structural and functional components
| Autor: | Michael Hammer, Kenneth I. Strauss, J. Kuźnicki, Lois Winsky, Jun Ichi Kawagoe, David M. Jacobowitz |
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| Rok vydání: | 1997 |
| Předmět: |
Recombinant Fusion Proteins
Molecular Sequence Data Restriction Mapping Nerve Tissue Proteins Biology Regulatory Sequences Nucleic Acid Transfection PC12 Cells Cellular and Molecular Neuroscience Exon Mice S100 Calcium Binding Protein G Genes Reporter Animals Humans Luciferase Cloning Molecular Luciferases Promoter Regions Genetic Molecular Biology Gene Cells Cultured Sequence Deletion Neurons Reporter gene Binding Sites Base Sequence Uterus Nucleic acid sequence Brain Promoter 3T3 Cells Molecular biology TATA Box Rats DNA-Binding Proteins nervous system Calbindin 2 Female Calretinin Sequence Alignment |
| Zdroj: | Brain research. Molecular brain research. 49(1-2) |
| ISSN: | 0169-328X |
| Popis: | The 5′ flanking region of the mouse calretinin gene was cloned and a 1.8 kbp region adjacent to exon 1 was sequenced. Putative upstream promoter and enhancer elements were identified, including appropriately positioned TATA and CAAT boxes (positions −50 and −68, respectively). There was considerable sequence and structural homology between mouse and human upstream elements. Neuron-restrictive activity was demonstrated via transfection of calretinin promoter-reporter constructs into primary embryonic mouse brain cultures expressing calretinin. In promoterless reporter constructs, the proximal upstream 1.5 kbp of the mouse calretinin gene boosted luciferase activity (up to 100-fold) exclusively in the neuronal population. Deletion analysis revealed the minimal promoter to be within the 95-bp proximal to the transcription start site. Transfections with SV40 promoter constructs in these cultures resulted in reporter gene expression predominantly in non-neuronal cells. Inserting the proximal 1.5 kbp of mouse calretinin upstream in SV40 promoter-reporter constructs reduced luciferase activity. Thus, calretinin upstream sequences increased reporter expression in cultured neurons and decreased expression from the SV40 promoter in non-neuronal cultured brain cells. The calretinin promoter contained relevant regulatory element consensus motifs and demonstrated in vitro neuron-restrictive bioactivity. |
| Databáze: | OpenAIRE |
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