کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
9141915 | 1163886 | 2005 | 11 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Transcriptional regulation of the murine C5a receptor gene: NF-Y is required for basal and LPS induced expression in macrophages and endothelial cells
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کلمات کلیدی
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
زیست شناسی مولکولی
پیش نمایش صفحه اول مقاله
![عکس صفحه اول مقاله: Transcriptional regulation of the murine C5a receptor gene: NF-Y is required for basal and LPS induced expression in macrophages and endothelial cells Transcriptional regulation of the murine C5a receptor gene: NF-Y is required for basal and LPS induced expression in macrophages and endothelial cells](/preview/png/9141915.png)
چکیده انگلیسی
The anaphylatoxin receptors of the complement system are important in immune defense but also play a role in autoimmune disease. Reports have demonstrated induced C5a receptor (C5aR) expression in a number of disease states, yet little is known about the regulation of this gene. We have examined sequences in the presumptive promoter-enhancer region in order to study the regulation of this gene. Rapid amplification of cDNA ends (RACE) analyses were used to identify the transcriptional start site, and we then cloned 2278Â bp of sequence from this region for use in luciferase assays. Deletion analyses of 5â² sequences demonstrated that the majority of this region is dispensable for expression in macrophages and endothelial cells (ECs). A 232Â bp region proximal to the transcription start site was fully capable of directing expression in macrophages and ECs, while being minimally active in cells that do not express the receptor. The transcriptional regulatory site most critical for this expression matches the consensus sequence for nuclear factor-Y (NF-Y) at position â96. Site-directed mutagenesis of this site resulted in a 70-90% decrease in luciferase activity depending on the cell type. Electrophoretic mobility shift/supershift assay (EMSA) analyses demonstrated the specific binding of NF-Y to labeled oligonucleotides containing the putative CCAAT site with macrophages and EC nuclear extracts, and antibodies to NF-Y were able to supershift this â96 NF-Y complex. We also demonstrate LPS leads to enhanced C5aR transcription and this is mediated predominantly through the NF-Y site. The data reported in this study might be critical for determination of transcription factors that can be targeted pharmacologically to modulate the expression of the C5aR in infectious disease or autoimmunity.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Molecular Immunology - Volume 42, Issue 11, July 2005, Pages 1405-1415
Journal: Molecular Immunology - Volume 42, Issue 11, July 2005, Pages 1405-1415
نویسندگان
Jason R. Hunt, Carol B. Martin, Brian K. Martin,