title: Definition of novel GP6 polymorphisms and major difference in haplotype frequencies between populations by a combination of in-depth exon resequencing and genotyping with tag single nucleotide polymorphisms.
abstract: BACKGROUND
  Common genetic variants of cell surface receptors contribute to differences in functional responses and disease susceptibility. We have previously shown that single nucleotide polymorphisms (SNPs) in platelet glycoprotein VI (GP6) determine the extent of response to agonist. In addition, SNPs in the GP6 gene have been proposed as risk factors for coronary artery disease.
  
  
  METHODS
  To completely characterize genetic variation in the GP6 gene we generated a high-resolution SNP map by sequencing the promoter, exons and consensus splice sequences in 94 non-related Caucasoids. In addition, we sequenced DNA encoding the ligand-binding domains of GP6 from non-human primates to determine the level of evolutionary conservation.
  
  
  RESULTS
  Eighteen SNPs were identified, six of which encoded amino acid substitutions in the mature form of the protein. The single non-synonymous SNP identified in the exons encoding the ligand-binding domains, encoding for a 103Leu > Val substitution, resulted in reduced ligand binding. Two common protein isoforms were confirmed in Caucasoid with frequencies of 0.82 and 0.15. Variation at the GP6 locus was characterized further by determining SNP frequency in over 2000 individuals from different ethnic backgrounds.
  
  
  CONCLUSIONS
  The SNPs were polymorphic in all populations studied although significant differences in allele frequencies were observed. Twelve additional GP6 protein isoforms were identified from the genotyping results and, despite extensive variation in GP6, the sequence of the ligand-binding domains is conserved. Sequences from non-human primates confirmed this observation. These data provide valuable information for the optimal selection of genetic variants for use in future association studies.
authors.affiliation: Department of Haematology, University of Cambridge and National Blood Service, Cambridge, UK. naw23@cam.ac.uk
authors.forename: N A
authors.initials: NA
authors.lastname: Watkins
authors.forename: M N
authors.initials: MN
authors.lastname: O'Connor
authors.forename: A
authors.initials: A
authors.lastname: Rankin
authors.forename: N
authors.initials: N
authors.lastname: Jennings
authors.forename: E
authors.initials: E
authors.lastname: Wilson
authors.forename: I J
authors.initials: IJ
authors.lastname: Harmer
authors.forename: L
authors.initials: L
authors.lastname: Davies
authors.forename: P A
authors.initials: PA
authors.lastname: Smethurst
authors.forename: F
authors.initials: F
authors.lastname: Dudbridge
authors.forename: R W
authors.initials: RW
authors.lastname: Farndale
authors.forename: W H
authors.initials: WH
authors.lastname: Ouwehand
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001792:Blood Platelets / Q000378:metabolism; D002352:Carrier Proteins / Q000378:metabolism; D002460:Cell Line; D004330:Drosophila / Q000235:genetics; D005091:Exons; D005787:Gene Frequency; D005838:Genotype; D006239:Haplotypes; D006801:Humans; D015810:Linkage Disequilibrium; D008969:Molecular Sequence Data; D010455:Peptides / Q000378:metabolism; D010980:Platelet Membrane Glycoproteins / Q000235:genetics* / Q000378:metabolism; D020641:Polymorphism, Single Nucleotide; D011323:Primates / Q000235:genetics; D011401:Promoter Regions, Genetic; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
chemical_list: D002352:Carrier Proteins; D010455:Peptides; D010980:Platelet Membrane Glycoproteins; D020033:Protein Isoforms; C435908:collagen-related peptide; C098341:platelet membrane glycoprotein VI