Abstract
Using transient expression of the E. coli Dam methylase gene and analysis of the distribution of methylated GATC sites, we studied the distribution of open chromatin regions within a 140 kb long human genome segment in HEK-293 cells. Dam methylated sites were found in gene introns, exons, and intergenic regions, and their distribution along DNA was uneven. There were regions of high and low density of Dam methylated GATC sites, presumably corresponding to “open” and “closed” chromatin regions, respectively, and to the functional profile of the genomic locus under study. The Dam methylation profile was also generally in agreement with transcriptional activity of genes in the locus. Moreover, DNA regions accessible to Dam methylase apparently coincided with those hypersensitive to DNase I.
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Acknowledgements
The authors thank E.A. Stukacheva and S.B. Akopov for carrying out transfections, V.K. Potapov and N.V. Skaptsova for oligonucleotide synthesis, T.L. Azhikina and T.V. Vinogradova for critical remarks and advises, V.V. Ruda for assistance at several steps of the work, Anne Olsen for cosmid clones and N.I. Medvedeva for preparation of cosmids. We thank B.O. Glotov for critical reading of the manuscript. The work was supported by the Scientific School program of the Russian Federation President (project NSh 2006.2003.4), and the program of the Russian Academy of Sciences on Molecular and Cellular Biology.
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Bulanenkova, S., Snezhkov, E., Nikolaev, L. et al. Identification and mapping of open chromatin regions within a 140 kb polygenic locus of human chromosome 19 using E. coli Dam methylase. Genetica 130, 83–92 (2007). https://doi.org/10.1007/s10709-006-0026-1
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DOI: https://doi.org/10.1007/s10709-006-0026-1