TY - JOUR
T1 - Functional characterization of promoter region polymorphisms of human CYP2C19 gene
AU - Rao, Uppugunduri Satyanarayana Chakradhara
AU - Devendran, Anichavezhi
AU - Satyamoorthy, Kapettu
AU - Shewade, Deepak Gopal
AU - Krishnamoorthy, Rajgopal
AU - Chandrasekaran, Adithan
PY - 2011/8
Y1 - 2011/8
N2 - CYP2C19 is an enzyme involved in the metabolism of several clinically important drugs. The variations in the CYP2C19 promoter region may alter the transcription of the gene by altering the interaction between the trans and cis-acting elements. In the present study, CYP2C19 promoter region with different variant alleles were cloned into a pGL-3 basic luciferase reporter vector and transfected into HepG2 cell lines. Subsequently, dual luciferase activity was measured to evaluate the activity of the promoter region. Gel shift assays with predicted binding sites for CCAAT displacement protein, activating transcription factor-2 and glucocorticoid receptor were performed. Results from this study indicate that few variations present in the putative cis-acting elements of the CYP2C19 promoter region such as -1442T>C, -779A>C and -98T>C -1498T>G and -828del>T alter the transcription of the gene. Specific binding with nuclear proteins was also observed in gel shift assays. This may account for the interindividual variations in gene expression and genotype dependant differences in gene transcription. The results also suggest the role of activating transcription factor-2 and CCAAT displacement repressor protein on CYP2C19 gene transcription.
AB - CYP2C19 is an enzyme involved in the metabolism of several clinically important drugs. The variations in the CYP2C19 promoter region may alter the transcription of the gene by altering the interaction between the trans and cis-acting elements. In the present study, CYP2C19 promoter region with different variant alleles were cloned into a pGL-3 basic luciferase reporter vector and transfected into HepG2 cell lines. Subsequently, dual luciferase activity was measured to evaluate the activity of the promoter region. Gel shift assays with predicted binding sites for CCAAT displacement protein, activating transcription factor-2 and glucocorticoid receptor were performed. Results from this study indicate that few variations present in the putative cis-acting elements of the CYP2C19 promoter region such as -1442T>C, -779A>C and -98T>C -1498T>G and -828del>T alter the transcription of the gene. Specific binding with nuclear proteins was also observed in gel shift assays. This may account for the interindividual variations in gene expression and genotype dependant differences in gene transcription. The results also suggest the role of activating transcription factor-2 and CCAAT displacement repressor protein on CYP2C19 gene transcription.
UR - http://www.scopus.com/inward/record.url?scp=80052483091&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=80052483091&partnerID=8YFLogxK
U2 - 10.1007/s11033-010-0537-9
DO - 10.1007/s11033-010-0537-9
M3 - Article
C2 - 21152987
AN - SCOPUS:80052483091
SN - 0301-4851
VL - 38
SP - 4171
EP - 4179
JO - Molecular Biology Reports
JF - Molecular Biology Reports
IS - 6
ER -