1. Dam methylase accessibility as an instrument for analysis of mammalian chromatin structure
- Author
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Lev G. Nikolaev, S. S. Bulanenkova, E. V. Snezhkov, Alena Kozlova, Eugene D. Sverdlov, S. B. Akopov, Tatyana L. Azhikina, and E. S. Kotova
- Subjects
Cancer Research ,Chromatin Immunoprecipitation ,Site-Specific DNA-Methyltransferase (Adenine-Specific) ,Transcription, Genetic ,Locus (genetics) ,Biology ,Transfection ,Electron Transport Complex IV ,Histones ,Epigenetics of physical exercise ,Dam methylase ,Escherichia coli ,Animals ,Humans ,Deoxyribonucleases, Type II Site-Specific ,Promoter Regions, Genetic ,Molecular Biology ,Genetics ,Mammals ,Genome, Human ,Promoter ,Acetylation ,Methylation ,DNA Methylation ,Chromatin Assembly and Disassembly ,Molecular biology ,Chromatin ,Housekeeping gene ,HEK293 Cells ,CpG site ,Genetic Loci ,DNA methylation ,CpG Islands ,Chromosomes, Human, Pair 19 ,Plasmids - Abstract
For a 140-kb human genome locus, an analysis of the distribution of Dam methylase accessible sites, DNase I sensitive and resistant regions, unmethylated CpG sites and acetylated histone H3 molecules was performed and compared with transcriptional activity of the genes within the locus. A direct correlation was found between the extent of Dam methylation and C5 cytosine (CpG) methylation. It was also demonstrated that promoter regions of all highly and moderately transcribed genes are highly accessible to methylation by Dam methylase. In contrast, promoters of non-transcribed genes showed a very low extent of Dam methylation. Promoter regions of non-transcribed genes were also highly CpG methylated, and the promoter and more distant 5'-regions of the housekeeping gene COX6B1 were substantially CpG-demethylated. Some highly Dam methylase accessible regions are present in the intergenic regions of the locus suggesting that the latter contain either unidentified non-coding transcripts or extended regulatory elements like locus control regions.
- Published
- 2011