PMID: 6165993Jan 1, 1981Paper

Amplified ribosomal RNA genes in a rat hepatoma cell line are enriched in 5-methylcytosine

Proceedings of the National Academy of Sciences of the United States of America
U TantravahiO J Miller

Abstract

In a rat hepatoma cell line, H4-IIE-C3, a 10-fold excess of 18S and 28S rRNA genes has been found in amplified chromosome regions. Antibodies to 5-methylcytidine bound extensively to the DNA of these regions, indicating a high level of DNA methylation. Most of the amplified rRNA genes were transcriptionally inactive, as shown by their failure to stain with silver. DNAs from the tumor cells and control rat hepatocytes grown with L-[methyl-14C]methionine were digested with restriction endonuclease EcoRI; the DNA fragments were separated by agarose gel electrophoresis, denatured, transferred to nitrocellulose filters, and hybridized to 32P-labeled rRNA or cDNA. Fragments containing the 18S or 28S rRNA coding sequences occurred in three major size classes; all three were rich in 5-methylcytosine. Analysis of EcoRI fragments of DNA from the tumor and control cells after digestion with Hpa II or Msp I endonuclease indicated that the 5'-C-C-G-G-3' sequences in most of the amplified rRNA genes were methylated. Analysis of the fragments produced by digestion with Hha I endonuclease indicated a high degree of methylation within its recognition sequence in the amplified rRNA genes as well. The association of hypermethylation with restrict...Continue Reading

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Citations

Jan 1, 1981·Human Genetics·U TantravahiO J Miller
Jan 1, 1983·Human Genetics·H G Schwarzacher, F Wachtler
Jan 1, 1983·Human Genetics·D N Cooper
Jan 1, 1992·Mammalian Genome : Official Journal of the International Mammalian Genome Society·K KawasakiN Shimizu
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