title: DNA methylation in the promoter of ribosomal RNA genes in human cells as determined by genomic sequencing.
abstract: Many RNA polymerase II- or III-transcribed genes are inactive when their promoter is methylated at critical CpG dinucleotides. We have applied the genomic sequencing method and a direct DNA blotting technique to analyze the extent of DNA methylation in the 5'-CpG-3' rich promoter region of the RNA polymerase I-transcribed ribosomal RNA genes (rDNA) in DNA from primary human cells, primary human tumor cells and human cell lines. In none of the analyzed primary human cells and primary human tumor cells was the DNA in the rDNA promoter region found to be detectably methylated. In contrast, in some of the cell lines this promoter is methylated in all 5'-CpG-3' dinucleotides in the majority of the approximately 200 ribosomal RNA gene copies. In actively growing cells, rDNA gene activity is a prerequisite for cell viability. The high levels of DNA methylation in the promoter region of rDNA in the human cell lines raise questions on the role of promoter methylation in these RNA polymerase I-transcribed genes. It is, however, conceivable that a subset of the about 200 rDNA copies per haploid genome have escaped methylation and account for the rRNA synthesis in these cell lines. Alternatively, complete 5'-CpG-3' promoter methylation may be compatible with promoter activity as demonstrated for certain viral genomes.
authors.affiliation: Division of Medical Genetics and Virology, Institute of Genetics, University of Cologne, Germany.
authors.forename: S
authors.initials: S
authors.lastname: Kochanek
authors.forename: K
authors.initials: K
authors.lastname: Hosokawa
authors.forename: G
authors.initials: G
authors.lastname: Schiedner
authors.forename: D
authors.initials: D
authors.lastname: Renz
authors.forename: W
authors.initials: W
authors.lastname: Doerfler
mesh_terms: D001483:Base Sequence; D002460:Cell Line; D002478:Cells, Cultured; D004247:DNA; D017931:DNA Primers; D004275:DNA, Ribosomal / Q000378:metabolism*; D003841:Deoxycytidine / Q000378:metabolism; D015226:Dinucleoside Phosphates / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D007624:KB Cells; D008745:Methylation; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D012318:RNA Polymerase I / Q000235:genetics; D012335:RNA, Ribosomal / Q000378:metabolism*; D014407:Tumor Cells, Cultured
chemical_list: D004247:DNA; D017931:DNA Primers; D004275:DNA, Ribosomal; D003841:Deoxycytidine; D015226:Dinucleoside Phosphates; D012318:RNA Polymerase I; D012335:RNA, Ribosomal; C015772:CpG dinucleotide