Identification and characterization of a novel 5 bp deletion in a putative insulin response element in the lipoprotein lipase gene
| Autor: | M. Ilyas Kamboh, Hamid Razzaghi, Li-Xia Yang, John E. Hokanson |
|---|---|
| Rok vydání: | 2009 |
| Předmět: |
Adult
Molecular Sequence Data Mutant Electrophoretic Mobility Shift Assay Plasma protein binding Biology Response Elements Gene Expression Regulation Enzymologic Article Cell Line Young Adult Exon Complementary DNA Chlorocebus aethiops Animals Humans Insulin Electrophoretic mobility shift assay Base Pairing Molecular Biology Gene Polymorphism Single-Stranded Conformational Aged Sequence Deletion ets-Domain Protein Elk-1 Lipoprotein lipase Base Sequence Reverse Transcriptase Polymerase Chain Reaction fungi Wild type DNA Exons Cell Biology Middle Aged Molecular biology Lipoprotein Lipase COS Cells Protein Binding |
| Zdroj: | Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids. 1791:1057-1065 |
| ISSN: | 1388-1981 |
| DOI: | 10.1016/j.bbalip.2009.06.003 |
| Popis: | Our aim was to identify an insulin response element (IRE) in the lipoprotein lipase (LPL) gene. We identified a 19 bp sequence as a putative IRE in LPL non-coding exon 10 using bioinformatics. Upon sequencing the IRE region, a novel 5 bp deletion was identified in Hispanics ( N = 406) with a carrier frequency of 4.2% but not in non-Hispanic whites ( N = 604) or Africans. Electrophoretic mobility shift assay revealed binding sites for regulatory factor(s) in muscle cell nuclear extracts with putative IRE sequence. Antibody supershift assay using human aorta smooth muscle cell nuclear extract revealed that Elk-1 specifically binds to putative IRE. TaqMan real-time RT-PCR of the 5 bp deletion, the mutant and wild type cDNA expressed in COS-1 and human muscle cells revealed that the 5 bp deletion was associated with modest reduction in LPL expression. There was also a slight reduction in LPL translation in the deletion mutant. Our data suggest the presence of an IRE in the 3′UTR of the LPL gene. |
| Databáze: | OpenAIRE |
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