Data from: Mechanisms of DNA methyltransferase 3A1-mediated DNA methylation of nucleosomes

DNA methyltransferase 3A1 (DNMT3A1) plays a crucial role in establishing
DNA methylation patterns that regulate gene expression and drive cellular
differentiation. In this study, we investigated the biochemical mechanisms
underlying DNMT3A1’s interactions with nucleosomes, the fundamental units
of chromatin. Using radiochemical activity assays, along with fluorescence
anisotropy and AlphaLISA binding assays, we demonstrated that DNMT3A1 has
multivalent interactions with nucleosomes, binding linker DNA and
nucleosome cores. Nanopore-based 5mC sequencing revealed that DNMT3A1
interactions with the nucleosome stimulate methylation of linker DNA up to
24 bp away from the nucleosome core. We suggest that the DNMT3A1 PWWP
domain interacts with distal linker DNA to facilitate uniform stimulation
of linker methylation. Additionally, our results indicate that DNMT3A1
binding is restricted to a single nucleosome, suggesting that its activity
is not allosterically regulated by neighboring nucleosomes. Together,
these findings provide new mechanistic insights into how DNMT3A1 engages
nucleosomes. This dataset includes unedited gel images of DNMT3A
purifications and nucleosome reconstitutions.