Genomic and phylogenetic footprinting at the epsilon-globin silencer region in intact human cells.
Chromatin structure at the silencer region of the epsilon-globin promoter was investigated using novel nitrogen mustards as probes of protein--DNA interactions. Sites of protection and enhancement that corresponded to known transcription factor binding sites were detected in both K562 and HeLa cells at this gene region. Protection was observed at several sites including the GATA-1/YY1 motifs. Of particular interest was a large 155 bp footprint that was observed at the epsilon-globin gene silencer region of the promoter. This large footprint was consistent with the presence of a positioned nucleosome core in intact human cells at this silencer region. Additionally, the DNA sequence at the epsilon-globin silencer and promoter was compared for 11 mammalian species. Significant areas of conservation were found that correlated with known transcription factor binding motifs. This phylogenetic footprinting analysis was compared to the genomic footprinting data at the epsilon-globin silencer region.
Temple
Mark D
MD
School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney NSW 2052, Australia.
Freebody
John
J
Murray
Vincent
V
D020816:Amino Acid Motifs; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002843:Chromatin / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D004249:DNA Damage; D005914:Globins / Q000235:genetics* / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D020014:K562 Cells; D008466:Mechlorethamine / Q000494:pharmacology; D008957:Models, Genetic; D008969:Molecular Sequence Data; D005810:Multigene Family; D009707:Nucleosomes / Q000378:metabolism; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D017422:Sequence Analysis, DNA; D013045:Species Specificity
D002843:Chromatin; D009707:Nucleosomes; D008466:Mechlorethamine; D005914:Globins; D004247:DNA