Structural context of disease-associated mutations and putative mechanism of autoinhibition revealed by X-Ray crystallographic analysis of the EZH2-SET domain

Stephen Antonysamy, Bradley Condon, Zhanna Druzina, Jeffrey B. Bonanno, Tarun Gheyi, Feiyu Zhang, Iain MacEwan, Aiping Zhang, Sheela Ashok, Logan Rodgers, Marijane Russell, John Gately Luz

Research output: Contribution to journalArticle

50 Citations (Scopus)

Abstract

The enhancer-of-zeste homolog 2 (EZH2) gene product is an 87 kDa polycomb group (PcG) protein containing a C-terminal methyltransferase SET domain. EZH2, along with binding partners, i.e., EED and SUZ12, upon which it is dependent for activity forms the core of the polycomb repressive complex 2 (PRC2). PRC2 regulates gene silencing by catalyzing the methylation of histone H3 at lysine 27. Both overexpression and mutation of EZH2 are associated with the incidence and aggressiveness of various cancers. The novel crystal structure of the SET domain was determined in order to understand disease-associated EZH2 mutations and derive an explanation for its inactivity independent of complex formation. The 2.00 Å crystal structure reveals that, in its uncomplexed form, the EZH2 C-terminus folds back into the active site blocking engagement with substrate. Furthermore, the S-adenosyl-L-methionine (SAM) binding pocket observed in the crystal structure of homologous SET domains is notably absent. This suggests that a conformational change in the EZH2 SET domain, dependent upon complex formation, must take place for cofactor and substrate binding activities to be recapitulated. In addition, the data provide a structural context for clinically significant mutations found in the EZH2 SET domain.

Original languageEnglish (US)
Article numbere84147
JournalPLoS One
Volume8
Issue number12
DOIs
StatePublished - Dec 19 2013

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Polycomb Repressive Complex 2
crystal structure
X-radiation
Crystal structure
X-Rays
mutation
X rays
Mutation
Genes
Polycomb-Group Proteins
S-Adenosylmethionine
Methylation
S-adenosylmethionine
methyltransferases
Methyltransferases
gene silencing
Substrates
histones
active sites
methylation

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Structural context of disease-associated mutations and putative mechanism of autoinhibition revealed by X-Ray crystallographic analysis of the EZH2-SET domain. / Antonysamy, Stephen; Condon, Bradley; Druzina, Zhanna; Bonanno, Jeffrey B.; Gheyi, Tarun; Zhang, Feiyu; MacEwan, Iain; Zhang, Aiping; Ashok, Sheela; Rodgers, Logan; Russell, Marijane; Luz, John Gately.

In: PLoS One, Vol. 8, No. 12, e84147, 19.12.2013.

Research output: Contribution to journalArticle

Antonysamy, S, Condon, B, Druzina, Z, Bonanno, JB, Gheyi, T, Zhang, F, MacEwan, I, Zhang, A, Ashok, S, Rodgers, L, Russell, M & Luz, JG 2013, 'Structural context of disease-associated mutations and putative mechanism of autoinhibition revealed by X-Ray crystallographic analysis of the EZH2-SET domain', PLoS One, vol. 8, no. 12, e84147. https://doi.org/10.1371/journal.pone.0084147
Antonysamy, Stephen ; Condon, Bradley ; Druzina, Zhanna ; Bonanno, Jeffrey B. ; Gheyi, Tarun ; Zhang, Feiyu ; MacEwan, Iain ; Zhang, Aiping ; Ashok, Sheela ; Rodgers, Logan ; Russell, Marijane ; Luz, John Gately. / Structural context of disease-associated mutations and putative mechanism of autoinhibition revealed by X-Ray crystallographic analysis of the EZH2-SET domain. In: PLoS One. 2013 ; Vol. 8, No. 12.
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