Graded methylation in the promoter and body of the O6-methylguanine DNA methyltransferase (MGMT) gene correlates with MGMT expression in human glioma cells

Joseph F. Costello, Bernard W Futscher, Keizo Tano, Dawn M. Graunke, Russell O. Pieper

Research output: Contribution to journalArticle

152 Citations (Scopus)

Abstract

Expression of the O6-methylguanine DNA methyl-transferase (MGMT) gene in human glioma cell lines is strongly associated with resistance to the chemotherapeutic agent 1,3-bis(2-chloroethyl)-1-nitrosourea. To examine the possibility that methylation of the body and promoter regions of the MGMT gene is associated with MGMT expression in a graded, rather than a completely on/off fashion, the present study analyzed the methylation status of the MGMT gene in human glioma cell lines exhibiting a wide range of MGMT expression. Methylation in the body of the gene was uniform within each cell line and correlated directly with MGMT expression. The level of MGMT promoter methylation was also graded across the cell lines, at 21 of 25 CpGs tested, but correlated inversely with MGMT expression. Two sites in the MGMT promoter were also much more accessible to restriction enzyme digestion, and thus in a more open chromatin conformation, in nuclei from high MGMT expressors relative to nuclei from cells with little or no MGMT expression. We conclude that the level of methylation, in both the body and promoter of the MGMT gene, is associated with MGMT expression in a graded fashion and may be important in setting the transcriptional state of the MGMT promoter through changes in chromatin structure.

Original languageEnglish (US)
Pages (from-to)17228-17237
Number of pages10
JournalJournal of Biological Chemistry
Volume269
Issue number25
StatePublished - Jun 24 1994

Fingerprint

Methylation
Methyltransferases
Glioma
Genes
DNA
Cells
Cell Line
O-(6)-methylguanine
Chromatin
Carmustine
Body Regions
Transferases
Cell Nucleus
Genetic Promoter Regions
Conformations
Digestion

ASJC Scopus subject areas

  • Biochemistry

Cite this

Graded methylation in the promoter and body of the O6-methylguanine DNA methyltransferase (MGMT) gene correlates with MGMT expression in human glioma cells. / Costello, Joseph F.; Futscher, Bernard W; Tano, Keizo; Graunke, Dawn M.; Pieper, Russell O.

In: Journal of Biological Chemistry, Vol. 269, No. 25, 24.06.1994, p. 17228-17237.

Research output: Contribution to journalArticle

@article{0c0f49ca24b74b4d84dcfdb705b6c3eb,
title = "Graded methylation in the promoter and body of the O6-methylguanine DNA methyltransferase (MGMT) gene correlates with MGMT expression in human glioma cells",
abstract = "Expression of the O6-methylguanine DNA methyl-transferase (MGMT) gene in human glioma cell lines is strongly associated with resistance to the chemotherapeutic agent 1,3-bis(2-chloroethyl)-1-nitrosourea. To examine the possibility that methylation of the body and promoter regions of the MGMT gene is associated with MGMT expression in a graded, rather than a completely on/off fashion, the present study analyzed the methylation status of the MGMT gene in human glioma cell lines exhibiting a wide range of MGMT expression. Methylation in the body of the gene was uniform within each cell line and correlated directly with MGMT expression. The level of MGMT promoter methylation was also graded across the cell lines, at 21 of 25 CpGs tested, but correlated inversely with MGMT expression. Two sites in the MGMT promoter were also much more accessible to restriction enzyme digestion, and thus in a more open chromatin conformation, in nuclei from high MGMT expressors relative to nuclei from cells with little or no MGMT expression. We conclude that the level of methylation, in both the body and promoter of the MGMT gene, is associated with MGMT expression in a graded fashion and may be important in setting the transcriptional state of the MGMT promoter through changes in chromatin structure.",
author = "Costello, {Joseph F.} and Futscher, {Bernard W} and Keizo Tano and Graunke, {Dawn M.} and Pieper, {Russell O.}",
year = "1994",
month = "6",
day = "24",
language = "English (US)",
volume = "269",
pages = "17228--17237",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "25",

}

TY - JOUR

T1 - Graded methylation in the promoter and body of the O6-methylguanine DNA methyltransferase (MGMT) gene correlates with MGMT expression in human glioma cells

AU - Costello, Joseph F.

AU - Futscher, Bernard W

AU - Tano, Keizo

AU - Graunke, Dawn M.

AU - Pieper, Russell O.

PY - 1994/6/24

Y1 - 1994/6/24

N2 - Expression of the O6-methylguanine DNA methyl-transferase (MGMT) gene in human glioma cell lines is strongly associated with resistance to the chemotherapeutic agent 1,3-bis(2-chloroethyl)-1-nitrosourea. To examine the possibility that methylation of the body and promoter regions of the MGMT gene is associated with MGMT expression in a graded, rather than a completely on/off fashion, the present study analyzed the methylation status of the MGMT gene in human glioma cell lines exhibiting a wide range of MGMT expression. Methylation in the body of the gene was uniform within each cell line and correlated directly with MGMT expression. The level of MGMT promoter methylation was also graded across the cell lines, at 21 of 25 CpGs tested, but correlated inversely with MGMT expression. Two sites in the MGMT promoter were also much more accessible to restriction enzyme digestion, and thus in a more open chromatin conformation, in nuclei from high MGMT expressors relative to nuclei from cells with little or no MGMT expression. We conclude that the level of methylation, in both the body and promoter of the MGMT gene, is associated with MGMT expression in a graded fashion and may be important in setting the transcriptional state of the MGMT promoter through changes in chromatin structure.

AB - Expression of the O6-methylguanine DNA methyl-transferase (MGMT) gene in human glioma cell lines is strongly associated with resistance to the chemotherapeutic agent 1,3-bis(2-chloroethyl)-1-nitrosourea. To examine the possibility that methylation of the body and promoter regions of the MGMT gene is associated with MGMT expression in a graded, rather than a completely on/off fashion, the present study analyzed the methylation status of the MGMT gene in human glioma cell lines exhibiting a wide range of MGMT expression. Methylation in the body of the gene was uniform within each cell line and correlated directly with MGMT expression. The level of MGMT promoter methylation was also graded across the cell lines, at 21 of 25 CpGs tested, but correlated inversely with MGMT expression. Two sites in the MGMT promoter were also much more accessible to restriction enzyme digestion, and thus in a more open chromatin conformation, in nuclei from high MGMT expressors relative to nuclei from cells with little or no MGMT expression. We conclude that the level of methylation, in both the body and promoter of the MGMT gene, is associated with MGMT expression in a graded fashion and may be important in setting the transcriptional state of the MGMT promoter through changes in chromatin structure.

UR - http://www.scopus.com/inward/record.url?scp=0028227862&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0028227862&partnerID=8YFLogxK

M3 - Article

C2 - 8006031

AN - SCOPUS:0028227862

VL - 269

SP - 17228

EP - 17237

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 25

ER -