Targeted methylation of CMV and E1A viral promoters

Chia Chen Hsu; Hsin Pai Li; Yu Hung Hung; Yu Wei Leu; Wu Hsiung Wu; Feng Sheng Wang; Kuan Der Lee; Pey Jium Chang; Chi Sheng Wu; Yen Jung Lu; Tim H.M. Huang; Yu Sun Chang; Shu Huei Hsiao

doi:10.1016/j.bbrc.2010.09.131

Targeted methylation of CMV and E1A viral promoters

Chia Chen Hsu, Hsin Pai Li, Yu Hung Hung, Yu Wei Leu, Wu Hsiung Wu, Feng Sheng Wang, Kuan Der Lee, Pey Jium Chang, Chi Sheng Wu, Yen Jung Lu, Tim H.M. Huang, Yu Sun Chang, Shu Huei Hsiao

Research output: Contribution to journal › Article › peer-review

62 Scopus citations

Abstract

DNA methylation is a gene-silencing and host defense system that can down-regulate viral gene expression in mammalian cells. An established targeted DNA methylation method was used to demonstrate that genome-integrated CMV and adenovirus type 5 E1A promoters were hypermethylated after MCF7 and HEK293 cells were transfected with in vitro methylated viral promoter fragments. In both cases, the targeted methylation-induced gene silencing could be reversed by addition of 5-aza-2'-deoxycytidine, confirming that the CMV and E1A promoters are regulated by DNA methylation. The kinetics of the targeted DNA methylation was determined using a reporter system in live cells. In conclusion, targeted DNA methylation is able to efficiently silence susceptible viral promoters and provides an alternative strategy to study the impact of loci-specific DNA methylation in viral gene expression.

Original language	English (US)
Pages (from-to)	228-234
Number of pages	7
Journal	Biochemical and Biophysical Research Communications
Volume	402
Issue number	2
DOIs	https://doi.org/10.1016/j.bbrc.2010.09.131
State	Published - Nov 12 2010
Externally published	Yes

Keywords

DNA methylation
Epigenetics
Targeted DNA methylation
Two-component EGFP reporter system
Viral promoter

ASJC Scopus subject areas

Molecular Biology
Biophysics
Biochemistry
Cell Biology

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10.1016/j.bbrc.2010.09.131

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title = "Targeted methylation of CMV and E1A viral promoters",

abstract = "DNA methylation is a gene-silencing and host defense system that can down-regulate viral gene expression in mammalian cells. An established targeted DNA methylation method was used to demonstrate that genome-integrated CMV and adenovirus type 5 E1A promoters were hypermethylated after MCF7 and HEK293 cells were transfected with in vitro methylated viral promoter fragments. In both cases, the targeted methylation-induced gene silencing could be reversed by addition of 5-aza-2'-deoxycytidine, confirming that the CMV and E1A promoters are regulated by DNA methylation. The kinetics of the targeted DNA methylation was determined using a reporter system in live cells. In conclusion, targeted DNA methylation is able to efficiently silence susceptible viral promoters and provides an alternative strategy to study the impact of loci-specific DNA methylation in viral gene expression.",

keywords = "DNA methylation, Epigenetics, Targeted DNA methylation, Two-component EGFP reporter system, Viral promoter",

author = "Hsu, {Chia Chen} and Li, {Hsin Pai} and Hung, {Yu Hung} and Leu, {Yu Wei} and Wu, {Wu Hsiung} and Wang, {Feng Sheng} and Lee, {Kuan Der} and Chang, {Pey Jium} and Wu, {Chi Sheng} and Lu, {Yen Jung} and Huang, {Tim H.M.} and Chang, {Yu Sun} and Hsiao, {Shu Huei}",

note = "Funding Information: This work was supported by the National Science Council in Taiwan ( NSC-96-2320-B-194-004 and NSC-95-2320-B-194-003 to S.H.H., NSC-98-3112-B-194-001 , NSC-97-2320-B-194-003-MY3 , and NSC-97-2627-B-006-003 to Y.W.L.). ",

year = "2010",

month = nov,

day = "12",

doi = "10.1016/j.bbrc.2010.09.131",

language = "English (US)",

volume = "402",

pages = "228--234",

journal = "Biochemical and Biophysical Research Communications",

issn = "0006-291X",

publisher = "Elsevier B.V.",

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TY - JOUR

T1 - Targeted methylation of CMV and E1A viral promoters

AU - Hsu, Chia Chen

AU - Li, Hsin Pai

AU - Hung, Yu Hung

AU - Leu, Yu Wei

AU - Wu, Wu Hsiung

AU - Wang, Feng Sheng

AU - Lee, Kuan Der

AU - Chang, Pey Jium

AU - Wu, Chi Sheng

AU - Lu, Yen Jung

AU - Huang, Tim H.M.

AU - Chang, Yu Sun

AU - Hsiao, Shu Huei

N1 - Funding Information: This work was supported by the National Science Council in Taiwan ( NSC-96-2320-B-194-004 and NSC-95-2320-B-194-003 to S.H.H., NSC-98-3112-B-194-001 , NSC-97-2320-B-194-003-MY3 , and NSC-97-2627-B-006-003 to Y.W.L.).

PY - 2010/11/12

Y1 - 2010/11/12

N2 - DNA methylation is a gene-silencing and host defense system that can down-regulate viral gene expression in mammalian cells. An established targeted DNA methylation method was used to demonstrate that genome-integrated CMV and adenovirus type 5 E1A promoters were hypermethylated after MCF7 and HEK293 cells were transfected with in vitro methylated viral promoter fragments. In both cases, the targeted methylation-induced gene silencing could be reversed by addition of 5-aza-2'-deoxycytidine, confirming that the CMV and E1A promoters are regulated by DNA methylation. The kinetics of the targeted DNA methylation was determined using a reporter system in live cells. In conclusion, targeted DNA methylation is able to efficiently silence susceptible viral promoters and provides an alternative strategy to study the impact of loci-specific DNA methylation in viral gene expression.

AB - DNA methylation is a gene-silencing and host defense system that can down-regulate viral gene expression in mammalian cells. An established targeted DNA methylation method was used to demonstrate that genome-integrated CMV and adenovirus type 5 E1A promoters were hypermethylated after MCF7 and HEK293 cells were transfected with in vitro methylated viral promoter fragments. In both cases, the targeted methylation-induced gene silencing could be reversed by addition of 5-aza-2'-deoxycytidine, confirming that the CMV and E1A promoters are regulated by DNA methylation. The kinetics of the targeted DNA methylation was determined using a reporter system in live cells. In conclusion, targeted DNA methylation is able to efficiently silence susceptible viral promoters and provides an alternative strategy to study the impact of loci-specific DNA methylation in viral gene expression.

KW - DNA methylation

KW - Epigenetics

KW - Targeted DNA methylation

KW - Two-component EGFP reporter system

KW - Viral promoter

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Targeted methylation of CMV and E1A viral promoters

Abstract

Keywords

ASJC Scopus subject areas

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