Regulatory genetic variation in the Flavin-containing monooxygenase 1 gene (FMO1): detection and functional consequences.
Doctoral thesis, UCL (University College London).
The amount of both FMO1 mRNA and protein in various human tissues has previously been shown to vary by as much as 10-fold between individuals. This is likely to be caused by genetic variation as the enzyme‟s expression profile is not affected by exogenous agents. DNA sequences controlling transcription (promoter regions) have been defined upstream of the three transcriptional start sites P0, P1 and P2 within different cell lines. The use of these promoters has also been defined in various mouse tissues by real-time PCR. A number of SNPs are present within the three defined FMO1 promoters in addition to the 3′UTR. Sequencing of individuals from the Corriel repository consisting of individuals from Africa, Asia and Europe has revealed further variation including a CT deletion at the P1 transcriptional start site. To test the effect of genetic variation high-throughput DNase I capillary footprinting has been used to check for the effect of SNPs on DNA-protein binding. The technique has been shown to detect the effects of mutations on DNA-protein binding but no differences have been seen for FMO1 SNPs within promoter regions as yet. The technique has the potential to rapidly analyse regulatory polymorphism in a number of genes and the effect in different tissues without the need for cloning or cell culture. A SNP which introduces an additional in-frame upstream translation initiation codon has been tested in-vitro for its effect on translational efficiency. The upstream ATG variation has been shown to have a 2-fold increase in protein expression over the downstream ATG and therefore individuals with this SNP are likely to produce 2-fold more FMO1 protein resulting in different effects to drugs.
|Title:||Regulatory genetic variation in the Flavin-containing monooxygenase 1 gene (FMO1): detection and functional consequences|
|Open access status:||An open access version is available from UCL Discovery|
|UCL classification:||UCL > School of Life and Medical Sciences > Faculty of Life Sciences > Biosciences (Division of) > Structural and Molecular Biology|
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