Functional characterization of promoter region polymorphisms of human CYP2C19 gene

Uppugunduri Satyanarayana Chakradhara Rao, Anichavezhi Devendran, Kapettu Satyamoorthy, Deepak Gopal Shewade, Rajgopal Krishnamoorthy, Adithan Chandrasekaran

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)4171-4179
Number of pages9
JournalMolecular Biology Reports
Volume38
Issue number6
DOIs
Publication statusPublished - 08-2011

Fingerprint

Genetic Promoter Regions
Activating Transcription Factor 2
Genes
Luciferases
Gels
Repressor Proteins
Glucocorticoid Receptors
Hep G2 Cells
Nuclear Proteins
Alleles
Binding Sites
Genotype
human CYP2C19 protein
Cytochrome P-450 CYP2C19
Gene Expression
Cell Line
Enzymes
Pharmaceutical Preparations
Proteins

All Science Journal Classification (ASJC) codes

  • Genetics
  • Molecular Biology

Cite this

Rao, U. S. C., Devendran, A., Satyamoorthy, K., Shewade, D. G., Krishnamoorthy, R., & Chandrasekaran, A. (2011). Functional characterization of promoter region polymorphisms of human CYP2C19 gene. Molecular Biology Reports, 38(6), 4171-4179. https://doi.org/10.1007/s11033-010-0537-9
Rao, Uppugunduri Satyanarayana Chakradhara ; Devendran, Anichavezhi ; Satyamoorthy, Kapettu ; Shewade, Deepak Gopal ; Krishnamoorthy, Rajgopal ; Chandrasekaran, Adithan. / Functional characterization of promoter region polymorphisms of human CYP2C19 gene. In: Molecular Biology Reports. 2011 ; Vol. 38, No. 6. pp. 4171-4179.
@article{3966052aa8b84fc481b0b5c613472006,
title = "Functional characterization of promoter region polymorphisms of human CYP2C19 gene",
abstract = "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.",
author = "Rao, {Uppugunduri Satyanarayana Chakradhara} and Anichavezhi Devendran and Kapettu Satyamoorthy and Shewade, {Deepak Gopal} and Rajgopal Krishnamoorthy and Adithan Chandrasekaran",
year = "2011",
month = "8",
doi = "10.1007/s11033-010-0537-9",
language = "English",
volume = "38",
pages = "4171--4179",
journal = "Molecular Biology Reports",
issn = "0301-4851",
publisher = "Springer Netherlands",
number = "6",

}

Rao, USC, Devendran, A, Satyamoorthy, K, Shewade, DG, Krishnamoorthy, R & Chandrasekaran, A 2011, 'Functional characterization of promoter region polymorphisms of human CYP2C19 gene', Molecular Biology Reports, vol. 38, no. 6, pp. 4171-4179. https://doi.org/10.1007/s11033-010-0537-9

Functional characterization of promoter region polymorphisms of human CYP2C19 gene. / Rao, Uppugunduri Satyanarayana Chakradhara; Devendran, Anichavezhi; Satyamoorthy, Kapettu; Shewade, Deepak Gopal; Krishnamoorthy, Rajgopal; Chandrasekaran, Adithan.

In: Molecular Biology Reports, Vol. 38, No. 6, 08.2011, p. 4171-4179.

Research output: Contribution to journalArticle

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

VL - 38

SP - 4171

EP - 4179

JO - Molecular Biology Reports

JF - Molecular Biology Reports

SN - 0301-4851

IS - 6

ER -