A 5′-Flanking C/G Pair at the Core Region Enhances the Recognition and Binding of Kaiso to Methylated DNA

• Published in: Journal of chemical information and modeling 2023-04, Vol.63 (7), p.2095-2103
• Main Authors: Thapa, Bidhya, Adhikari, Narayan P., Tiwari, Purushottam B., Chapagain, Prem P.
• Format: Article
• Language: English
• Subjects: Binding; Computational Biochemistry; CpG Islands; DNA - chemistry; DNA Methylation; Gene Expression Regulation; Gene sequencing; Molecular dynamics; Nucleotides; Protein Binding; Proteins; Recognition; Transcription Factors - chemistry; Zinc Fingers - genetics
• ISSN: 1549-9596; 1549-960X
• DOI: 10.1021/acs.jcim.2c01294

Methyl CpG binding proteins (MBPs) are transcription factors that recognize the methylated CpG sites in DNA and mediate the DNA methylation signal into various downstream cellular processes. The C2H2 zinc finger (ZF) protein, Kaiso, also an MBP, preferentially binds to two symmetrically methylated CpG sites in DNA sequences via C-terminal C2H2 ZF domains and mediates the transcription regulation process. Investigation of the molecular mechanism of the recognition of methylated DNA (meDNA) by Kaiso is important to understand how this protein reads and translates this methylation signal into downstream transcription outcomes. Despite previous studies in Kaiso-meDNA interactions, detailed structural investigations on the sequence-specific interaction of Kaiso with the meDNA sequence are still lacking. In this work, we used molecular modeling and molecular dynamics (MD) simulation-based computational approaches to investigate the recognition of various methylated DNA sequences by Kaiso. Our MD simulation results show that the Kaiso-meDNA interaction is sequence specific. The recognition of meDNA by Kaiso is enhanced in the MeECad sequence compared to the MeCG2 sequence. Compared to the 5′-flanking T/A pair in MeCG2, both MeCG2_mutCG and MeECad sequences show that a C/G base pair allows GLU535 of Kaiso to preferably recognize and bind the core mCpG site. The core mCGmCG site is crucial for the recognition process and formation of a stable complex. Our results reveal that the 5′-flanking nucleotides are also important for the enhanced binding and recognition of methylated sites.