Cell type-specific activation of the cytomegalovirus promoter by dimethylsulfoxide and 5-Aza-2′-deoxycytidine

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Abstract

The cytomegalovirus promoter is a very potent promoter commonly used for driving the expression of transgenes, though it gradually becomes silenced in stably transfected cells. We examined the methylation status of the cytomegalovirus promoter in two different cell lines and characterized its mechanisms of activation by dimethylsulfoxide and 5-Aza-2′-deoxycytidine. The cytomegalovirus promoter stably transfected into Chinese hamster ovary cells is suppressed by DNA methylation-independent mechanisms, which is different from the rat embryonic cardiomyoblast H9c2-Fluc.3 cells in which the cytomegalovirus promoter is silenced by methylation. Dimethylsulfoxide and 5-Aza-2′-deoxycytidine can activate the cytomegalovirus promoter in both cell types by overlapping mechanisms. Dimethylsulfoxide activates the cytomegalovirus promoter in Chinese hamster ovary cells by promoting histone acetylation and the activation of p38 mitogen-activated protein kinase and nuclear factor κB (NFκB) signaling pathways, while 5-Aza-2′-deoxycytidine increases histone acetylation and activates the nuclear factor κB but not the p38 mitogen-activated protein kinase pathway. In H9c2-Fluc.3 cells, both agents promote demethylation of the cytomegalovirus promoter, and enhance its activity exclusively through activation of the nuclear factor κB pathway and to a lesser extent of the p38 mitogen-activated protein kinase pathway. Our findings suggest that suppression and activation of the cytomegalovirus promoter are cell type-specific. These results may be used for developing strategies to enhance the expression of transgenes and the production of recombinant proteins encoded by transgenes controlled by a cytomegalovirus promoter.

Original languageEnglish (US)
Pages (from-to)1944-1955
Number of pages12
JournalInternational Journal of Biochemistry and Cell Biology
Volume40
Issue number9
DOIs
StatePublished - Mar 10 2008

Fingerprint

decitabine
Dimethyl Sulfoxide
Cytomegalovirus
p38 Mitogen-Activated Protein Kinases
Chemical activation
Acetylation
Methylation
Cells
Histones
Transgenes
Recombinant Proteins
Cricetulus
Rats
Ovary
DNA Methylation

Keywords

  • 5-Aza-2′-deoxycytidine
  • Cytomegalovirus promoter
  • Dimethylsulfoxide
  • NFκB
  • p38 MAP kinase

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology

Cite this

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title = "Cell type-specific activation of the cytomegalovirus promoter by dimethylsulfoxide and 5-Aza-2′-deoxycytidine",
abstract = "The cytomegalovirus promoter is a very potent promoter commonly used for driving the expression of transgenes, though it gradually becomes silenced in stably transfected cells. We examined the methylation status of the cytomegalovirus promoter in two different cell lines and characterized its mechanisms of activation by dimethylsulfoxide and 5-Aza-2′-deoxycytidine. The cytomegalovirus promoter stably transfected into Chinese hamster ovary cells is suppressed by DNA methylation-independent mechanisms, which is different from the rat embryonic cardiomyoblast H9c2-Fluc.3 cells in which the cytomegalovirus promoter is silenced by methylation. Dimethylsulfoxide and 5-Aza-2′-deoxycytidine can activate the cytomegalovirus promoter in both cell types by overlapping mechanisms. Dimethylsulfoxide activates the cytomegalovirus promoter in Chinese hamster ovary cells by promoting histone acetylation and the activation of p38 mitogen-activated protein kinase and nuclear factor κB (NFκB) signaling pathways, while 5-Aza-2′-deoxycytidine increases histone acetylation and activates the nuclear factor κB but not the p38 mitogen-activated protein kinase pathway. In H9c2-Fluc.3 cells, both agents promote demethylation of the cytomegalovirus promoter, and enhance its activity exclusively through activation of the nuclear factor κB pathway and to a lesser extent of the p38 mitogen-activated protein kinase pathway. Our findings suggest that suppression and activation of the cytomegalovirus promoter are cell type-specific. These results may be used for developing strategies to enhance the expression of transgenes and the production of recombinant proteins encoded by transgenes controlled by a cytomegalovirus promoter.",
keywords = "5-Aza-2′-deoxycytidine, Cytomegalovirus promoter, Dimethylsulfoxide, NFκB, p38 MAP kinase",
author = "Prakash Radhakrishnan and Basma, {Hesham E} and Klinkebiel, {David L} and Judith Christman and Pi-Wan Cheng",
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T1 - Cell type-specific activation of the cytomegalovirus promoter by dimethylsulfoxide and 5-Aza-2′-deoxycytidine

AU - Radhakrishnan, Prakash

AU - Basma, Hesham E

AU - Klinkebiel, David L

AU - Christman, Judith

AU - Cheng, Pi-Wan

PY - 2008/3/10

Y1 - 2008/3/10

N2 - The cytomegalovirus promoter is a very potent promoter commonly used for driving the expression of transgenes, though it gradually becomes silenced in stably transfected cells. We examined the methylation status of the cytomegalovirus promoter in two different cell lines and characterized its mechanisms of activation by dimethylsulfoxide and 5-Aza-2′-deoxycytidine. The cytomegalovirus promoter stably transfected into Chinese hamster ovary cells is suppressed by DNA methylation-independent mechanisms, which is different from the rat embryonic cardiomyoblast H9c2-Fluc.3 cells in which the cytomegalovirus promoter is silenced by methylation. Dimethylsulfoxide and 5-Aza-2′-deoxycytidine can activate the cytomegalovirus promoter in both cell types by overlapping mechanisms. Dimethylsulfoxide activates the cytomegalovirus promoter in Chinese hamster ovary cells by promoting histone acetylation and the activation of p38 mitogen-activated protein kinase and nuclear factor κB (NFκB) signaling pathways, while 5-Aza-2′-deoxycytidine increases histone acetylation and activates the nuclear factor κB but not the p38 mitogen-activated protein kinase pathway. In H9c2-Fluc.3 cells, both agents promote demethylation of the cytomegalovirus promoter, and enhance its activity exclusively through activation of the nuclear factor κB pathway and to a lesser extent of the p38 mitogen-activated protein kinase pathway. Our findings suggest that suppression and activation of the cytomegalovirus promoter are cell type-specific. These results may be used for developing strategies to enhance the expression of transgenes and the production of recombinant proteins encoded by transgenes controlled by a cytomegalovirus promoter.

AB - The cytomegalovirus promoter is a very potent promoter commonly used for driving the expression of transgenes, though it gradually becomes silenced in stably transfected cells. We examined the methylation status of the cytomegalovirus promoter in two different cell lines and characterized its mechanisms of activation by dimethylsulfoxide and 5-Aza-2′-deoxycytidine. The cytomegalovirus promoter stably transfected into Chinese hamster ovary cells is suppressed by DNA methylation-independent mechanisms, which is different from the rat embryonic cardiomyoblast H9c2-Fluc.3 cells in which the cytomegalovirus promoter is silenced by methylation. Dimethylsulfoxide and 5-Aza-2′-deoxycytidine can activate the cytomegalovirus promoter in both cell types by overlapping mechanisms. Dimethylsulfoxide activates the cytomegalovirus promoter in Chinese hamster ovary cells by promoting histone acetylation and the activation of p38 mitogen-activated protein kinase and nuclear factor κB (NFκB) signaling pathways, while 5-Aza-2′-deoxycytidine increases histone acetylation and activates the nuclear factor κB but not the p38 mitogen-activated protein kinase pathway. In H9c2-Fluc.3 cells, both agents promote demethylation of the cytomegalovirus promoter, and enhance its activity exclusively through activation of the nuclear factor κB pathway and to a lesser extent of the p38 mitogen-activated protein kinase pathway. Our findings suggest that suppression and activation of the cytomegalovirus promoter are cell type-specific. These results may be used for developing strategies to enhance the expression of transgenes and the production of recombinant proteins encoded by transgenes controlled by a cytomegalovirus promoter.

KW - 5-Aza-2′-deoxycytidine

KW - Cytomegalovirus promoter

KW - Dimethylsulfoxide

KW - NFκB

KW - p38 MAP kinase

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VL - 40

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EP - 1955

JO - International Journal of Biochemistry and Cell Biology

JF - International Journal of Biochemistry and Cell Biology

SN - 1357-2725

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