title: Functional Validation of a Common Nonsynonymous Coding Variant in ZC3HC1 Associated With Protection From Coronary Artery Disease.
abstract: BACKGROUND
  Although virtually all coronary artery disease associated single-nucleotide polymorphisms identified by genome-wide association studies (GWAS) are in noncoding regions of the genome, a common polymorphism in ZC3HC1 (rs11556924), resulting in an arginine (Arg) to histidine (His) substitution in its encoded protein, NIPA (Nuclear Interacting Partner of Anaplastic Lyphoma Kinase) is linked to a protection from coronary artery disease. NIPA plays a role in cell cycle progression, but the functional consequences of this polymorphism have not been established.
  
  
  RESULTS
  Here we demonstrate that total ZC3HC1 expression in whole blood is similar across genotypes, despite expression being slightly biased toward the risk allele in heterozygotes. At the protein level, the protective His363 NIPA variant exhibits increased phosphorylation of a critical serine residue (Ser354) and higher protein expression as compared with the Arg363 variant. Binding experiments indicate that neither SKP1 (S-phase kinase-associated protein 1) nor CCNB1 binding were affected by the polymorphism. Despite similar nuclear distribution, NIPA His363 exhibits greater nuclear mobility. NIPA suppression results in a modest reduction of proliferation in vascular smooth muscle cells, but given low proliferative capacity, a significant effect of the variant was not noted. By contrast, we demonstrate that the protective variant reduces cell proliferation in HeLa cells.
  
  
  CONCLUSIONS
  These findings extend the genetic association between rs11556924 and coronary artery disease risk by characterizing its effects on the encoded protein, NIPA. The resulting amino acid change Arg363His is associated with increased expression and nuclear mobility, as well as lower rates of cell growth in HeLa cells, further supporting a role for cell proliferation in atherosclerosis and its clinical consequences.
authors.affiliation: From the Atherogenomics Laboratory, University of Ottawa Heart Institute, Canada.
authors.forename: Tara
authors.initials: T
authors.lastname: Linseman
authors.affiliation: From the Atherogenomics Laboratory, University of Ottawa Heart Institute, Canada. rmcpherson@ottawaheart.ca ssoubeyrand@ottawaheart.ca.
authors.forename: Sébastien
authors.initials: S
authors.lastname: Soubeyrand
authors.affiliation: From the Atherogenomics Laboratory, University of Ottawa Heart Institute, Canada.
authors.forename: Amy
authors.initials: A
authors.lastname: Martinuk
authors.affiliation: From the Atherogenomics Laboratory, University of Ottawa Heart Institute, Canada.
authors.forename: Majid
authors.initials: M
authors.lastname: Nikpay
authors.affiliation: From the Atherogenomics Laboratory, University of Ottawa Heart Institute, Canada.
authors.forename: Paulina
authors.initials: P
authors.lastname: Lau
authors.affiliation: From the Atherogenomics Laboratory, University of Ottawa Heart Institute, Canada. rmcpherson@ottawaheart.ca ssoubeyrand@ottawaheart.ca.
authors.forename: Ruth
authors.initials: R
authors.lastname: McPherson
mesh_terms: D021581:Active Transport, Cell Nucleus; D048868:Adaptor Proteins, Signal Transducing / Q000097:blood / Q000235:genetics*; D018797:Cell Cycle Proteins / Q000097:blood / Q000235:genetics*; D002467:Cell Nucleus / Q000378:metabolism; D049109:Cell Proliferation; D003324:Coronary Artery Disease / Q000097:blood / Q000175:diagnosis / Q000235:genetics* / Q000517:prevention & control*; D056744:Cyclin B1 / Q000378:metabolism; D004843:Epistasis, Genetic; D005819:Genetic Markers; D020022:Genetic Predisposition to Disease; D057809:HEK293 Cells; D006367:HeLa Cells; D061307:Human Umbilical Vein Endothelial Cells / Q000378:metabolism; D006801:Humans; D015810:Linkage Disequilibrium; D009131:Muscle, Smooth, Vascular / Q000378:metabolism; D032389:Myocytes, Smooth Muscle / Q000378:metabolism; D009687:Nuclear Proteins / Q000097:blood / Q000235:genetics*; D010641:Phenotype; D010766:Phosphorylation; D020641:Polymorphism, Single Nucleotide; D065840:Protective Factors; D018570:Risk Assessment; D012307:Risk Factors; D014162:Transfection
chemical_list: D048868:Adaptor Proteins, Signal Transducing; C534699:CCNB1 protein, human; D018797:Cell Cycle Proteins; D056744:Cyclin B1; D005819:Genetic Markers; D009687:Nuclear Proteins; C477288:ZC3HC1 protein, human
keywords: CCNB1; NIPA; ZC3HC1; atherogenesis; coronary artery disease; genome-wide association study; genomics; phosphorylation