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Binding of human antigen R (HuR) to an AU-rich element (ARE) in the 3′untranslated region (3′UTR) reduces the expression of decay accelerating factor (DAF)
- Source :
- Molecular Immunology. 47:2545-2551
- Publication Year :
- 2010
- Publisher :
- Elsevier BV, 2010.
-
Abstract
- We have investigated the role of the 3'untranslated region (3'UTR) in the expression of decay accelerating factor (DAF), one of the major membrane regulators of Complement activation. We show here that the 3'UTR of DAF contains an adenylate uridine rich element (ARE) AUUUAUUUAUAUUUAUUUA, which belongs to Class II Cluster 4 of the AU-rich element-containing mRNA (ARED) database. Enhanced Green Fluorescent Protein (EGFP) Reporter constructs containing the DAF 3'UTR showed reduced levels of expression when transfected into a variety of cell lines compared to 3'UTR reporter constructs without the ARE sequence. Furthermore, the inhibitor of mRNA transcription Actinomycin D had a much stronger effect on mRNA half-life of the ARE-containing 3'UTR demonstrating that this ARE destabilises the mRNA. Electrophoretic Mobility Shift Assays (EMSA) using biotinylated RNA probes, demonstrated that cytoplasmic Human antigen R (HuR) bound to the DAF ARE. Transfection experiments using HuR specific siRNA increased DAF expression whilst plasmids containing the coding sequence of HuR had the opposite effect, demonstrating that HuR reduced the stability of DAF mRNA and suggesting that it is of importance in regulating the expression of DAF. These data suggest that modulators of HuR could potentially be used to alter DAF expression and therefore increase the susceptibility of malignant cells to immunotherapy.
- Subjects :
- Immunology
Biology
Cell Line
ELAV-Like Protein 1
Green fluorescent protein
Transcription (biology)
Humans
Coding region
RNA, Small Interfering
3' Untranslated Regions
Molecular Biology
Decay-accelerating factor
AU-rich element
Messenger RNA
Base Sequence
CD55 Antigens
Three prime untranslated region
fungi
RNA-Binding Proteins
Transfection
Molecular biology
ELAV Proteins
Gene Expression Regulation
Antigens, Surface
Dactinomycin
Protein Binding
Subjects
Details
- ISSN :
- 01615890
- Volume :
- 47
- Database :
- OpenAIRE
- Journal :
- Molecular Immunology
- Accession number :
- edsair.doi.dedup.....762381fca2c8f79ff648a296ec59cc1f