title: Multicapillary gel electrophoresis based analysis of genetic variants in the WFS1 gene.
abstract: The WFS1 gene is one of the thoroughly investigated targets in diabetes research, variants of the gene were suggested to be the genetic components of the common forms (type 1 and type 2) of diabetes. Our project focused on the analysis of polymorphisms (rs4689388, rs148797429, rs4273545) localized in the WFS1 promoter region. Although submarine gel electrophoresis based approaches were also employed in the genetic tests, it was demonstrated that multicapillary electrophoresis offers a state of the art approach for reliable high-throughput SNP and VNTR analysis. Association studies were carried out in a case-control setup. Luciferase reporter assay was employed to test the effect of the investigated loci on the activity of gene expression in vitro. Significant association could be demonstrated between all three polymorphisms and type 2 diabetes in both allele- and genotype-wise settings even using Bonferroni correction. It is notable; however, that the three loci were in strong linkage disequilibrium, thus the observed associations cannot be considered as separate effects. Molecular analyses showed that the rs4273545 GT SNP played a role in the regulation of transcription in vitro. However, this effect took place only in the presence of the region including the rs148797429 site, although this latter locus did not have its own impact on the regulation of gene expression. The paper provides genotyping protocols readily applicable in any multiplex SNP and VNTR analyses, moreover confirms and extends previous results about the role of WFS1 polymorphisms in the genetic risk of diabetes mellitus.
authors.affiliation: Department of Medical Chemistry, Molecular Biology and Pathobiochemistry, Semmelweis University, Budapest, Hungary.
authors.forename: Zsuzsanna
authors.initials: Z
authors.lastname: Elek
authors.affiliation: Department of Medical Chemistry, Molecular Biology and Pathobiochemistry, Semmelweis University, Budapest, Hungary.
authors.forename: Réka
authors.initials: R
authors.lastname: Dénes
authors.affiliation: Department of Medical Chemistry, Molecular Biology and Pathobiochemistry, Semmelweis University, Budapest, Hungary.
authors.forename: Susanne
authors.initials: S
authors.lastname: Prokop
authors.affiliation: The 2nd Department of Internal Medicine, Semmelweis University, Budapest, Hungary.
authors.forename: Anikó
authors.initials: A
authors.lastname: Somogyi
authors.affiliation: SCIEX Separations, Brea, CA, USA.
authors.forename: Handy
authors.initials: H
authors.lastname: Yowanto
authors.affiliation: SCIEX Separations, Brea, CA, USA.
authors.forename: Jane
authors.initials: J
authors.lastname: Luo
authors.affiliation: SCIEX Separations, Darmstadt, Germany.
authors.forename: Manfred
authors.initials: M
authors.lastname: Souquet
authors.affiliation: SCIEX Separations, Brea, CA, USA.|MTA-PE Translational Glycomics Group, University of Pannonia, Veszprém, Hungary.|Horváth Laboratory of Bioseparation Sciences, University of Debrecen, Debrecen, Hungary.
authors.forename: András
authors.initials: A
authors.lastname: Guttman
authors.affiliation: Department of Medical Chemistry, Molecular Biology and Pathobiochemistry, Semmelweis University, Budapest, Hungary. ronai.zsolt@med.semmelweis-univ.hu.
authors.forename: Zsolt
authors.initials: Z
authors.lastname: Rónai
mesh_terms: D000328:Adult; D000368:Aged; D004587:Electrophoresis, Agar Gel; D019075:Electrophoresis, Capillary / Q000379:methods*; D005260:Female; D055106:Genome-Wide Association Study; D006801:Humans; D015810:Linkage Disequilibrium; D008297:Male; D008565:Membrane Proteins / Q000235:genetics*; D008875:Middle Aged; D018598:Minisatellite Repeats; D020641:Polymorphism, Single Nucleotide; D011401:Promoter Regions, Genetic
chemical_list: D008565:Membrane Proteins; C114987:wolframin protein
keywords: Capillary gel electrophoresis; Diabetes mellitus; SNP; Single base primer extension; WFS1