title: Novel photodynamic effect of a psoralen-conjugated oligonucleotide for the discrimination of the methylation of cytosine in DNA.
abstract: DNA methylation and demethylation significantly affect the deactivation and activation processes of gene expression significantly. In particular, C-5-methylation of cytosine in the CpG islands is important for the epigenetic modification in genes, which plays a key role in regulating gene expression. The determination of the location and frequency of DNA methylation is important for the elucidation of the mechanisms of cell differentiation and carcinogenesis. Here we designed a psoralen-conjugated oligonucleotide (PS-oligo) for the discrimination of 5-methylcytosine (5-mC) in DNA. The cross-linking behavior of psoralen derivatives with pyrimidine bases, such as thymine, uracil and cytosine has been well discussed, but there are no reports which have examined whether cross-linking efficiency of psoralen with cytosine would be changed with or without C-5 methylation. We found that the cross-linking efficiency of PS-oligo with target-DNA containing 5-mC was greatly increased compared to the case of target-DNA without 5-mC, approximately seven-fold higher. Here we report a new aspect of the photocross-linking behavior of psoralen with 5-mC that is applicable to a simple, sequence-specific and quantitative analysis for the discrimination of 5-mC in DNA, which can be applicable to study the epigenetic behavior of gene expressions.
authors.affiliation: Department of Biomolecular Engineering, Kyoto Institute of Technology, Matsugasaki, Sakuo-ku, Kyoto, Japan.
authors.forename: Asako
authors.initials: A
authors.lastname: Yamayoshi
authors.forename: Yohei
authors.initials: Y
authors.lastname: Matsuyama
authors.forename: Mikihiko
authors.initials: M
authors.lastname: Kushida
authors.forename: Akio
authors.initials: A
authors.lastname: Kobori
authors.forename: Akira
authors.initials: A
authors.lastname: Murakami
mesh_terms: D003596:Cytosine / Q000737:chemistry*; D019175:DNA Methylation; D011564:Furocoumarins / Q000737:chemistry*; D009841:Oligonucleotides / Q000737:chemistry*; D010778:Photochemotherapy
chemical_list: D011564:Furocoumarins; D009841:Oligonucleotides; D003596:Cytosine