Beta3 integrin haplotype influences gene regulation and plasma von Willebrand factor activity.
The Leu33Pro polymorphism of the gene encoding beta(3) integrin (ITGB3) is associated with acute coronary syndromes and influences platelet aggregation. Three common promoter polymorphisms have also been identified. The aims of this study were to (1) investigate the influence of the ITGB3 -400C/A, -425A/C and -468G/A promoter polymorphisms on reporter gene expression and nuclear protein binding and (2) determine genotype and haplotype associations with platelet alpha(IIb)beta(3) receptor density. Promoter haplotypes were introduced into an ITGB3 promoter-pGL3 construct by site directed mutagenesis and luciferase reporter gene expression analysed in HEL and HMEC-1 cells. Binding of nuclear proteins was assessed by electrophoretic mobility shift assay. The association of ITGB3 haplotype with platelet alpha(IIb)beta(3) receptor density was determined in 223 subjects. Species conserved motifs were identified in the ITGB3 promoter in the vicinity of the three polymorphisms. The GAA, GCC, AAC, AAA and ACC constructs induced approximately 50% increased luciferase expression relative to the GAC construct in both cell types. Haplotype analysis including Leu33Pro indicated five common haplotypes; no associations between ITGB3 haplotypes and receptor density were found. However, the GCC-Pro33 haplotype was associated with significantly higher vWF activity (128.6 [112.1-145.1]%) compared with all other haplotypes (107.1 [101.2-113.0]%, p=0.02). In conclusion, the GCC-Pro33 haplotype was associated with increased vWF activity but not with platelet alpha(IIb)beta(3) receptor density, which may indicate ITGB3 haplotype influences endothelial function.
Academic Unit of Molecular Vascular Medicine, Leeds Institute for Genetics, Health and Therapeutics, University of Leeds, UK.
Katie E
KE
Payne
Paul F
PF
Bray
Peter J
PJ
Grant
Angela M
AM
Carter
D000328:Adult; D000818:Animals; D001483:Base Sequence; D019295:Computational Biology; D017124:Conserved Sequence; D005260:Female; D005340:Fibrinogen / Q000032:analysis; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D006239:Haplotypes; D006801:Humans; D039661:Integrin beta3 / Q000737:chemistry / Q000235:genetics*; D008156:Luciferases / Q000235:genetics; D008297:Male; D051379:Mice; D008875:Middle Aged; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000378:metabolism; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000235:genetics; D011110:Polymorphism, Genetic; D011392:Proline / Q000737:chemistry / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D014841:von Willebrand Factor / Q000235:genetics* / Q000378:metabolism*
C061248:ITGB3 protein, human; D039661:Integrin beta3; D009687:Nuclear Proteins; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D014841:von Willebrand Factor; D005340:Fibrinogen; D011392:Proline; D008156:Luciferases