Abstract
Histone lysine methylation has a pivotal role in regulating the chromatin. Histone modifiers, including histone methyl transferases (HMTases), have clear roles in human carcinogenesis but the extent of their functions and regulation are not well understood. The NSD family of HMTases comprised of three members (NSD1, NSD2/MMSET/WHSC1, and NSD3/WHSC1L) are oncogenes aberrantly expressed in several cancers, suggesting their potential to serve as novel therapeutic targets. However, the substrate specificity of the NSDs and the molecular mechanism of histones H3 and H4 recognition and methylation have not yet been established. Herein, we investigated the in vitro mechanisms of histones H3 and H4 recognition and modifications by the catalytic domain of NSD family members. In this study, we quantified in vitro mono-, di- and tri- methylations on H3K4, H3K9, H3K27, H3K36, H3K79, and H4K20 by the carboxyl terminal domain (CTD) of NSD1, NSD2 and NSD3, using histone as substrate. Next, we used a molecular modelling approach and docked 6-mer peptides H3K4 a.a. 1-7; H3K9 a.a. 5-11; H3K27 a.a. 23-29; H3K36 a.a. 32-38; H3K79 a.a. 75-81; H4K20 a.a. 16-22 with the catalytic domain of the NSDs to provide insight into lysine-marks recognition and methylation on histones H3 and H4. Our data highlight the versatility of NSD1, NSD2, and NSD3 for recognizing and methylating several histone lysine marks on histones H3 and H4. Our work provides a basis to design selective and specific NSDs inhibitors. We discuss the relevance of our findings for the development of NSD inhibitors amenable for novel chemotherapies.
Highlights
Histone lysine methylation has a pivotal role in regulating the chromatin
The carboxyl terminal domain (CTD) of nuclear receptor-binding SET domain (NSD) members can methylate multiple histone lysines in vitro To examine whether the CTD of NSDs: Met1999 (NSD1), NSD2, and NSD3 recognizes and methylates multiple lysines of H3 and H4 in vitro, we quantified the panmethylation of NSD1-CTD, NSD2-CTD, and NSD3CTD on H3K4, H3K9, H3K27, H3K36, H3K79, and H4K20 (Figure 2)
NSD1-CTD, NSD2-CTD, and NSD3-CTD exhibit similar substrate recognition and methylation properties, but reduced methylation activities were observed for NSD3-CTD (Figure 2)
Summary
Histone lysine methylation has a pivotal role in regulating the chromatin. Histone modifiers, including histone methyl transferases (HMTases), have clear roles in human carcinogenesis but the extent of their functions and regulation are not well understood. The substrate specificity of the NSDs and the molecular mechanism of histones H3 and H4 recognition and methylation have not yet been established. Covalent histone modifications are key in chromatin regulatory mechanisms. One such histone modification, lysine methylation, can have both activating and repressive functions on transcriptional events. Histone lysine methyltransferases (HMTases) are transcriptional coregulators that target specific lysines on histones H3 and H4, and can transfer up to three methyl groups (Kme, Kme, and Kme3) [1].
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