title: Multidrug resistance polypeptide 1 (MDR1, ABCB1) variant 3435C>T affects mRNA stability.
abstract: OBJECTIVE
  ABCB1 (multidrug resistance 1 polypeptide, MDR1, Pgp) is a multispecific efflux transporter of drugs and xenobiotics. Among numerous polymorphisms in human ABCB1, the synonymous SNP 3435C > T has been associated with decreased mRNA and protein levels, via unknown mechanisms.
  
  
  METHODS
  To search for cis-acting polymorphism affecting transcription or mRNA processing, we used 3435C > T as a marker single nucleotide polymorphism (SNP), for measuring differences in allelic mRNA expression. Ratios of allelic abundance in genomic DNA and mRNA (after conversion to cDNA) were measured quantitatively with a primer extension assay, in human liver samples.
  
  
  RESULTS
  mRNA expression of the 3435C allele was significantly higher than that of the 3435T allele (3435C/3435T ratios ranging from 1.06-1.61). Cotransfection of equal amounts of ABCB1 expression plasmids containing 3435C or 3435T also revealed higher 3435C mRNA expression. Increasing 3435C/3435T ratios after cessation of transcription indicated that the 3435C > T substitution decreases mRNA stability. 3435C > T is in strong linkage disequilibrium with two other coding SNPs (1236C > T and 2677G > T) forming two abundant haplotypes (ABCB1*1 and ABCB1*13). Transfection of all possible combinations of these three SNPs demonstrated that only 3435T is associated with lower mRNA levels. Calculations of mRNA folding, using Mfold, suggested an effect on mRNA secondary structure.
  
  
  CONCLUSIONS
  the abundant 3435C > T SNP appears to be a main factor in allelic variation of ABCB1 mRNA expression in the liver, by changing mRNA stability.
authors.affiliation: Department of Pharmacology, Program in Pharmacogenomics, College of Medicine and Public Health, The Ohio State University, Columbus, Ohio 43210, USA.
authors.forename: Danxin
authors.initials: D
authors.lastname: Wang
authors.forename: Andrew D
authors.initials: AD
authors.lastname: Johnson
authors.forename: Audrey C
authors.initials: AC
authors.lastname: Papp
authors.forename: Deanna L
authors.initials: DL
authors.lastname: Kroetz
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Sadée
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000235:genetics*; D000483:Alleles; D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D004247:DNA / Q000032:analysis; D019450:Genes, MDR / Q000235:genetics*; D005838:Genotype; D006801:Humans; D008099:Liver / Q000502:physiology*; D009690:Nucleic Acid Conformation; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D020871:RNA Stability; D014162:Transfection
chemical_list: D018528:ATP-Binding Cassette Transporters; D004247:DNA