Tag Archives: COG7

Background and seeks Coronary artery disease (CAD) risk is associated with

Background and seeks Coronary artery disease (CAD) risk is associated with non-coding genetic variants at the phosphatase and actin regulating protein 1(gene encodes an actin-binding protein with phosphatase regulating activity. we GS-1101 demonstrate that PHACTR1 is expressed as multiple previously uncharacterized transcripts in macrophages foam cells GS-1101 lymphocytes and endothelial cells. Immunoblotting confirmed a total absence of PHACTR1 in vascular smooth muscle cells. Real-time quantitative PCR showed that is regulated by atherogenic and inflammatory stimuli. In aortic endothelial cells oxLDL and TNF-alpha both upregulated an intermediate length transcript. A short transcript expressed only in immune cells was upregulated in macrophages by oxidized low-density lipoprotein and oxidized phospholipids but suppressed by lipopolysaccharide or TNF-alpha. In primary human macrophages we identified a novel expression quantitative trait locus (eQTL) specific for this short transcript whereby the risk allele at CAD risk SNP rs9349379 is associated with reduced PHACTR1 expression similar to the effect of an inflammatory stimulus. Conclusions Our data GS-1101 demonstrate that is a key atherosclerosis candidate gene since it is regulated by atherogenic COG7 stimuli in macrophages and endothelial cells and we identify an effect of the genetic risk variant on PHACTR1 expression in macrophages that is similar GS-1101 to that of an inflammatory stimulus. locus are associated with the specific phenotypes of early onset myocardial infarction coronary artery calcification [11] [12] and with an intermediate phenotype of impaired central hemodynamic indices indicating abnormal vascular stiffness [13]. The locus is pleiotropic since the protective alleles of the CAD risk SNPs are associated with an increased risk of ischemic stroke caused by cervical artery dissection a form of non-atherosclerotic vascular disease [14]. The variants reported in these studies lie in an intronic region of over 250?kb away from any other gene. A genetic fine mapping study suggested that rs9349379 was the most likely causative variant at the locus and it was associated with expression of mRNA in composite right coronary artery tissue [15]. An expression quantitative trait locus (eQTL) study assaying gene expression in diverse human tissues showed that rs9349379 also affected mRNA expression in aortic artery and tibial artery tissue [16]. In a whole genome epigenetic and expression study we recently exhibited that is one of the most highly upregulated genes in macrophages exposed to oxLDL [17]. The family consists of 4 genes encoding proteins that interact directly with both actin and protein phosphatase 1 (PP1) [18]. was originally cloned from a rat brain cDNA library using a yeast two-hybrid system with PP1 as bait [18]. The human gene is usually on chromosome 6 and a transcript with a 1743?bp open reading frame has been cloned from human brain [19]. The resulting 580 amino acid protein has 4 highly conserved actin-binding RPEL domains and both mouse and rat orthologues have been shown to bind actin [18] [20]. Human PHACTR1 protein contains nuclear localization signal (NLS) motifs at both ends of the protein that in the mouse ortholog have been shown to facilitate importin-dependent nuclear translocation in response to serum stimulation [20]. Binding of the RPEL domains to G-actin maintains PHACTR1 in a cytoplasmic location [20]. Serum induces GS-1101 Rho-dependent remodeling of actin from G- to F-actin and translocation of PHACTR1 into the nucleus [20]. PP1 is usually a part of a family of serine/threonine phosphatases that are present in both the nucleus and cytoplasm [21]. The enzymatic specificity of PP1 enzymes is usually achieved in part by their association with accessory proteins [22]. PHACTR1 may function as an accessory protein since PHACTR1 binds to PP1 and inhibits its activity gene is usually forecasted to encode multiple transcripts which have not really been characterized. Within this research we hypothesized that PHACTR1 is certainly governed by atherogenic or inflammatory stimuli which its appearance is certainly inspired by CAD-associated hereditary variations. After building the appearance of PHACTR1 proteins in atherosclerotic lesions as well as the profile of individual transcripts in major cell types involved with atherosclerosis we motivated the responses of the transcripts to inflammatory stimuli also to atherogenic lipid. The function of PHACTR1 in CAD was highlighted by its great quantity in macrophages and foam cells in individual atherosclerotic plaque. The CAD risk allele at SNP rs9349379 was connected with.