TY - JOUR
T1 - Identification of three new cis-regulatory IRF5 polymorphisms
T2 - In vitro studies
AU - Alonso-Perez, Elisa
AU - Fernandez-Poceiro, Romina
AU - Lalonde, Emilie
AU - Kwan, Tony
AU - Calaza, Manuel
AU - Gomez-Reino, Juan J.
AU - Majewski, Jacek
AU - Gonzalez, Antonio
N1 - Funding Information:
We thank Carmen Pena-Pena for her expert technical assistance. EAP has been the recipient of an Instituto de Salud Carlos III predoctoral bursary. This project was supported by grants PI08/0744 and PI11/01048 and by a RETICS Program grant (RD08/0075 (RIER)) from the Instituto de Salud Carlos III (Spain), which are partially financed by the European Regional Development Fund program of the European Union.
PY - 2013/8/13
Y1 - 2013/8/13
N2 - Background: Polymorphisms in the interferon regulatory factor 5 (IRF5) gene are associated with susceptibility to systemic lupus erythematosus, rheumatoid arthritis and other diseases through independent risk and protective haplotypes. Several functional polymorphisms are already known, but they do not account for the protective haplotypes that are tagged by the minor allele of rs729302.Methods: Polymorphisms in linkage disequilibrium (LD) with rs729302 or particularly associated with IRF5 expression were selected for functional screening, which involved electrophoretic mobility shift assays (EMSAs) and reporter gene assays.Results: A total of 54 single-nucleotide polymorphisms in the 5' region of IRF5 were genotyped. Twenty-four of them were selected for functional screening because of their high LD with rs729302 or protective haplotypes. In addition, two polymorphisms were selected for their prominent association with IRF5 expression. Seven of these twenty-six polymorphisms showed reproducible allele differences in EMSA. The seven were subsequently analyzed in gene reporter assays, and three of them showed significant differences between their two alleles: rs729302, rs13245639 and rs11269962. Haplotypes including the cis-regulatory polymorphisms correlated very well with IRF5 mRNA expression in an analysis based on previous data.Conclusion: We have found that three polymorphisms in LD with the protective haplotypes of IRF5 have differential allele effects in EMSA and in reporter gene assays. Identification of these cis-regulatory polymorphisms will allow more accurate analysis of transcriptional regulation of IRF5 expression, more powerful genetic association studies and deeper insight into the role of IRF5 in disease susceptibility.
AB - Background: Polymorphisms in the interferon regulatory factor 5 (IRF5) gene are associated with susceptibility to systemic lupus erythematosus, rheumatoid arthritis and other diseases through independent risk and protective haplotypes. Several functional polymorphisms are already known, but they do not account for the protective haplotypes that are tagged by the minor allele of rs729302.Methods: Polymorphisms in linkage disequilibrium (LD) with rs729302 or particularly associated with IRF5 expression were selected for functional screening, which involved electrophoretic mobility shift assays (EMSAs) and reporter gene assays.Results: A total of 54 single-nucleotide polymorphisms in the 5' region of IRF5 were genotyped. Twenty-four of them were selected for functional screening because of their high LD with rs729302 or protective haplotypes. In addition, two polymorphisms were selected for their prominent association with IRF5 expression. Seven of these twenty-six polymorphisms showed reproducible allele differences in EMSA. The seven were subsequently analyzed in gene reporter assays, and three of them showed significant differences between their two alleles: rs729302, rs13245639 and rs11269962. Haplotypes including the cis-regulatory polymorphisms correlated very well with IRF5 mRNA expression in an analysis based on previous data.Conclusion: We have found that three polymorphisms in LD with the protective haplotypes of IRF5 have differential allele effects in EMSA and in reporter gene assays. Identification of these cis-regulatory polymorphisms will allow more accurate analysis of transcriptional regulation of IRF5 expression, more powerful genetic association studies and deeper insight into the role of IRF5 in disease susceptibility.
UR - http://www.scopus.com/inward/record.url?scp=84881301951&partnerID=8YFLogxK
U2 - 10.1186/ar4262
DO - 10.1186/ar4262
M3 - ???researchoutput.researchoutputtypes.contributiontojournal.article???
C2 - 23941291
AN - SCOPUS:84881301951
SN - 1478-6354
VL - 15
JO - Arthritis Research and Therapy
JF - Arthritis Research and Therapy
IS - 4
M1 - R82
ER -