Thermal Stability Changes in Telomeric G-Quadruplex Structures Due to <i>N</i>⁶-Methyladenosine Modification

• Main Authors: Ryohei Wada, Wataru Yoshida
• Format: Article
• Language: English
• Published: MDPI AG 2021-02-01
• Series: Epigenomes
• Subjects: G-quadruplex; telomere; <i>N</i>⁶-methyladenosine; thermal stability
• Online Access: https://www.mdpi.com/2075-4655/5/1/5

<i>N</i>⁶-methyladenosine modification (m⁶dA) has recently been identified in eukaryote genomic DNA. The methylation destabilizes the duplex structure when the adenine forms a Watson–Crick base pair, whereas the methylation on a terminal unpaired adenine stabilizes the duplex structure by increasing the stacking interaction. In this study, the effects of m⁶dA modification on the thermal stability of four distinct telomeric G-quadruplex (G4) structures were investigated. The m⁶dA-modified telomeric oligonucleotide d[AGGG(TTAGGG)₃] that forms a basket-type G4 in Na⁺, d[(TTAGGG)₄TT] that forms a hybrid-type G4 in K⁺ (Form-2), d[AAAGGG(TTAGGG)₃AA] that forms a hybrid-type G4 in K⁺ (Form-1), and d[GGG(TTAGGG)₃T] that forms a basket-type G4 with two G-tetrads in K⁺ (Form-3) were analyzed. Circular dichroism melting analysis demonstrated that (1) A7- and A19-methylation destabilized the basket-type G4 structure that formed in Na⁺, whereas A13-methylation stabilized the structure; (2) A15-methylation stabilized the Form-2 G4 structure; (3) A15- and A21-methylations stabilized the Form-1 G4 structure; and (4) A12-methylation stabilized the Form-3 G4 structure. These results suggest that m⁶dA modifications may affect the thermal stability of human telomeric G4 structures in regulating the biological functions.