title: Analysis of human glutathione S-transferase alpha 1 (hGSTA1) gene promoter polymorphism using denaturing high performance liquid chromatography (DHPLC).
abstract: OBJECTIVE
  The GST enzyme, encoded by hGSTA1 gene, catalyses the GSH dependant detoxification of a variety of carcinogenic metabolites and alkylating chemotherapeutic agents. Two genetic variants of hGSTA1, namely hGSTA1*A and hGSTA1*B, are characterized by three linked SNPs, of which -52 G>A variation being solely responsible for the differential promoter activity of hGSTA1. Individuals homozygous for hGSTA1*B have low hepatic expression of hGSTA1. Given the time and labor consuming PCR-RFLP method and the direct prediction of -52 G>A variation, we opted to establish a high throughput DHPLC procedure for the characterization of hGSTA1 variants.
  
  
  METHODS
  117 DNA samples from South India were included in the study. Control samples were generated for DHPLC using conventional PCR-RFLP technique. Heteroduplexes were produced by in vitro mixing of control DNA samples (hGSTA1*A) to all the samples which are subsequently subjected to DHPLC analysis. The samples were analyzed for the presence of heteroduplexes from the chromatographic profiles.
  
  
  CONCLUSIONS
  From the total of 117 samples, 43.5% are homozygous for hGSTA1*A allele, 13% are homozygous for hGSTA1*B allele and 43.5% are hGSTA1*A/B heterozygotes. This is, to our knowledge, the first report on the use of DHPLC for the evaluation of hGSTA1 gene promoter polymorphism.
authors.affiliation: Department of Pharmacology, PRIST University, Puducherry 605007, India. koumar@gmail.com
authors.forename: K
authors.initials: K
authors.lastname: Koumaravelou
authors.forename: Z
authors.initials: Z
authors.lastname: Shoaib
authors.forename: C
authors.initials: C
authors.lastname: Adithan
authors.forename: D
authors.initials: D
authors.lastname: Charron
authors.forename: A
authors.initials: A
authors.lastname: Srivastava
authors.forename: R
authors.initials: R
authors.lastname: Tamouza
authors.forename: R
authors.initials: R
authors.lastname: Krishnamoorthy
mesh_terms: D002851:Chromatography, High Pressure Liquid; D005982:Glutathione Transferase / Q000235:genetics*; D006801:Humans; D016133:Polymerase Chain Reaction; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012680:Sensitivity and Specificity
chemical_list: C484555:GSTA1 protein, human; D005982:Glutathione Transferase