title: In vitro histone lysine methylation by NSD1, NSD2/MMSET/WHSC1 and NSD3/WHSC1L.
abstract: BACKGROUND
  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.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Kyungpook National University, School of Applied Biosciences, Life Sciences and Agriculture building #3, room 309, 80 Daehak-ro, Daegu, Buk-gu, 702-701, Republic of Korea. morimori088@gmail.com.
authors.forename: Masayo
authors.initials: M
authors.lastname: Morishita
authors.affiliation: Kyungpook National University, School of Applied Biosciences, Life Sciences and Agriculture building #3, room 309, 80 Daehak-ro, Daegu, Buk-gu, 702-701, Republic of Korea. daanmevius@gmail.com.
authors.forename: Damiaan
authors.initials: D
authors.lastname: Mevius
authors.affiliation: Kyungpook National University, School of Applied Biosciences, Life Sciences and Agriculture building #3, room 309, 80 Daehak-ro, Daegu, Buk-gu, 702-701, Republic of Korea. eric.diluccio@gmail.com.
authors.forename: Eric
authors.initials: E
authors.lastname: di Luccio
mesh_terms: D020134:Catalytic Domain; D000076983:Histone Methyltransferases; D011495:Histone-Lysine N-Methyltransferase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006657:Histones / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques / Q000379:methods*; D047908:Intracellular Signaling Peptides and Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D008239:Lysine / Q000378:metabolism; D008745:Methylation; D008958:Models, Molecular; D009687:Nuclear Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D017433:Protein Structure, Secondary; D012097:Repressor Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D006657:Histones; D047908:Intracellular Signaling Peptides and Proteins; D009687:Nuclear Proteins; D012097:Repressor Proteins; D000076983:Histone Methyltransferases; D011495:Histone-Lysine N-Methyltransferase; C113272:NSD1 protein, human; C419586:NSD2 protein, human; C432968:NSD3 protein, human; D008239:Lysine