title: Transcriptional regulation of the human FPR2/ALX gene: evidence of a heritable genetic variant that impairs promoter activity.
abstract: Lipoxin (LX) A(4,) a main endogenous stop-signal of inflammation, activates the G-protein-coupled receptor FPR2/ALX, which triggers potent anti-inflammatory signaling in vivo. Thus, the regulation of FPR2/ALX expression may have pathophysiological and therapeutic relevance. Here, we mapped a nucleotide sequence with strong FPR2/ALX promoter activity. Chromatin immunoprecipitation revealed specificity protein 1 (Sp1) binding to the core promoter. Site-directed mutagenesis of the Sp1 cis-acting element and Sp1 overexpression established that this transcription factor is key for maximal promoter activity, which is instead suppressed by DNA methylation. LXA(4) enhanced FPR2/ALX promoter activity (+74%) and mRNA expression (+87.5%) in MDA-MB231 cells. A single nucleotide mutation (A/G) was detected in the core promoter of one subject with history of cardiovascular disease and of his two daughters. This mutation reduced by ∼35-90% the promoter activity in vitro. Moreover, neutrophils from individuals carrying the A/G variant displayed ∼10- and 3-fold reduction in FPR2/ALX mRNA and protein, respectively, compared with cells from their relatives or healthy volunteers expressing the wild-type allele. These results uncover FPR2/ALX transcriptional regulation and provide the first evidence of mutations that affect FPR2/ALX transcription, thus opening new opportunities for the understanding of the LXA(4)-FPR2/ALX axis in human disease.
authors: Department of Biomedical Sciences, Center of Excellence on Aging, Ce.S.I., G. d'Annunzio University Foundation, Via Colle dell'Ara, 66013 Chieti, Italy.
authors: Felice
authors: F
authors: Simiele
authors: Antonio
authors: A
authors: Recchiuti
authors: Domenico
authors: D
authors: Mattoscio
authors: Antonella
authors: A
authors: De Luca
authors: Eleonora
authors: E
authors: Cianci
authors: Sara
authors: S
authors: Franchi
authors: Valentina
authors: V
authors: Gatta
authors: Alessandro
authors: A
authors: Parolari
authors: Josè Pablo
authors: JP
authors: Werba
authors: Marina
authors: M
authors: Camera
authors: Bartolo
authors: B
authors: Favaloro
authors: Mario
authors: M
authors: Romano
mesh_terms: D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D015153:Blotting, Western; D002460:Cell Line; D045744:Cell Line, Tumor; D019175:DNA Methylation; D005786:Gene Expression Regulation; D014644:Genetic Variation; D006801:Humans; D008957:Models, Genetic; D008969:Molecular Sequence Data; D020641:Polymorphism, Single Nucleotide; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding; D044042:Receptors, Formyl Peptide / Q000235:genetics* / Q000378:metabolism; D044067:Receptors, Lipoxin / Q000235:genetics* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016329:Sp1 Transcription Factor / Q000378:metabolism; D024363:Transcription Initiation Site; D014158:Transcription, Genetic / Q000235:genetics*
chemical_list: C105101:FPR2 protein, human; D044042:Receptors, Formyl Peptide; D044067:Receptors, Lipoxin; D016329:Sp1 Transcription Factor