Histone and DNA binding ability studies of the NSD subfamily of PWWP domains

The NSD proteins, namely NSD1, NSD2 and NSD3, are lysine methyltransferases, which catalyze mono and di-methylation of histone H3K36. They are multi-domain proteins, including two PWWP domains (PWWP1 and PWWP2) separated by some other domains. These proteins act as potent oncoproteins and are implicated in various cancers. However the biological functions of these PWWP domains are still largely unknown. To better understand the functions of these proteins' PWWP domains, we cloned, expressed and purified all the PWWP domains of these NSD proteins to characterize their interactions with methylated histone peptides and dsDNA by quantitative binding assays and crystallographic analysis. Our studies indicate that all these PWWP domains except NSD1_PWWP1 bind to trimethylated H3K36, H3K79 peptides and dsDNA weakly. Our crystal structures uncover that the NDS3_PWWP2 and NSD2_PWWP1 domains, which hold an extremely long alpha-helix and alpha-helix bundle, respectively, need a conformation adjustment to interact with nucleosome. (C) 2021 Elsevier Inc. All rights reserved.

Automated summary

NSD proteins, including NSD1, NSD2, and NSD3, are lysine methyltransferases that catalyze mono and di-methylation of histone H3K36. The PWWP domains within these proteins weakly bind to trimethylated H3K36, H3K79 peptides, and dsDNA, except for NSD1_PWWP1. Structural analysis reveals that NSD3_PWWP2 and NSD2_PWWP1 domains require conformation adjustments to interact with nucleosomes.