title: Characterization of a human 12/15-lipoxygenase promoter variant associated with atherosclerosis identifies vimentin as a promoter binding protein.
abstract: BACKGROUND
  Sequence variation in the human 12/15 lipoxygenase (ALOX15) has been associated with atherosclerotic disease. We functionally characterized an ALOX15 promoter polymorphism, rs2255888, previously associated with carotid plaque burden.
  
  
  RESULTS
  We demonstrate specific in vitro and in vivo binding of the cytoskeletal protein, vimentin, to the ALOX15 promoter. We show that the two promoter haplotypes carrying alternate alleles at rs2255888 exhibit significant differences in promoter activity by luciferase reporter assay in two cell lines. Differences in in-vitro vimentin-binding to and formation of DNA secondary structures in the polymorphic promoter sequence are also detected by electrophoretic mobility shift assay and biophysical analysis, respectively. We show regulation of ALOX15 protein by vimentin.
  
  
  CONCLUSIONS
  This study suggests that vimentin binds the ALOX15 promoter and regulates its promoter activity and protein expression. Sequence variation that results in changes in DNA conformation and vimentin binding to the promoter may be relevant to ALOX15 gene regulation.
authors.affiliation: Research Center for Human Genetics, Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center at Houston, Houston, Texas, United States of America. Susmita.Samanta@bcm.edu
authors.forename: Susmita
authors.initials: S
authors.lastname: Samanta
authors.forename: Kurtis
authors.initials: K
authors.lastname: Anderson
authors.forename: Sean
authors.initials: S
authors.lastname: Moran
authors.forename: David
authors.initials: D
authors.lastname: Hawke
authors.forename: David
authors.initials: D
authors.lastname: Gorenstein
authors.forename: Myriam
authors.initials: M
authors.lastname: Fornage
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001093:Arachidonate 15-Lipoxygenase / Q000235:genetics*; D050197:Atherosclerosis / Q000201:enzymology* / Q000235:genetics*; D001483:Base Sequence; D018076:DNA, Complementary / Q000235:genetics; D020022:Genetic Predisposition to Disease; D006239:Haplotypes / Q000235:genetics; D006801:Humans; D008156:Luciferases / Q000378:metabolism; D061986:MCF-7 Cells; D051379:Mice; D008969:Molecular Sequence Data; D041681:NIH 3T3 Cells; D009690:Nucleic Acid Conformation; D009841:Oligonucleotides / Q000235:genetics; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D011401:Promoter Regions, Genetic; D011485:Protein Binding / Q000187:drug effects / Q000235:genetics; D012492:Salts / Q000494:pharmacology; D014162:Transfection; D014746:Vimentin / Q000378:metabolism*
chemical_list: D018076:DNA, Complementary; D009841:Oligonucleotides; D012492:Salts; D014746:Vimentin; C515716:ALOX15 protein, human; D001093:Arachidonate 15-Lipoxygenase; D008156:Luciferases