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Research Article

DNA methylation patterns associated with asparagine synthetase expression in asparagine-overproducing and -auxotrophic cells.

I L Andrulis, M T Barrett
I L Andrulis
Research Institute, Mt. Sinai Hospital, Ontario, Canada.
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M T Barrett
Research Institute, Mt. Sinai Hospital, Ontario, Canada.
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DOI: 10.1128/MCB.9.7.2922
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ABSTRACT

In Chinese hamster ovary cells, the gene for asparagine synthetase, which spans 20 kilobase pairs, was found to contain a cluster of potential sites for CpG methylation in a 1-kilobase-pair region surrounding the first exon. Fourteen of the sites that could be assayed for methylation by MspI-HpaII digestions were found in this region, with an additional nine MspI sites spread throughout the remainder of the gene. The methylation status of the gene was analyzed in a series of cell lines that differed in the amount of asparagine synthetase activity. The level of expression showed a direct correlation with the extent of methylation of a subset of the MspI sites found in the 5' region of the gene. The rest of the gene was completely methylated in most cell lines. Wild-type cells, which expressed a basal level of asparagine synthetase activity, were partially demethylated in the 5' region. In contrast, asparagine-requiring N3 cells, which lacked detectable mRNA for asparagine synthetase, were methylated throughout the entire gene. Spontaneous revertants of strain N3, selected for growth in asparagine-free medium, exhibited extensive hypomethylation of the asparagine synthetase gene. The methylation pattern of the gene in cell lines that overproduced the enzyme was also examined. Albizziin-resistant cell lines, which had amplified copies of the gene, were extensively demethylated in the 5' region. Overexpression of asparagine synthetase in beta-aspartyl hydroxamate-resistant lines without amplified copies of the gene was also correlated with DNA hypomethylation.

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DNA methylation patterns associated with asparagine synthetase expression in asparagine-overproducing and -auxotrophic cells.
I L Andrulis, M T Barrett
Molecular and Cellular Biology Jul 1989, 9 (7) 2922-2927; DOI: 10.1128/MCB.9.7.2922

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DNA methylation patterns associated with asparagine synthetase expression in asparagine-overproducing and -auxotrophic cells.
I L Andrulis, M T Barrett
Molecular and Cellular Biology Jul 1989, 9 (7) 2922-2927; DOI: 10.1128/MCB.9.7.2922
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