TY - JOUR
T1 - Structural context of disease-associated mutations and putative mechanism of autoinhibition revealed by X-Ray crystallographic analysis of the EZH2-SET domain
AU - Antonysamy, Stephen
AU - Condon, Bradley
AU - Druzina, Zhanna
AU - Bonanno, Jeffrey B.
AU - Gheyi, Tarun
AU - Zhang, Feiyu
AU - MacEwan, Iain
AU - Zhang, Aiping
AU - Ashok, Sheela
AU - Rodgers, Logan
AU - Russell, Marijane
AU - Luz, John Gately
N1 - Funding Information:
We thank Matthieu Shapira and colleagues for discussions and for sharing their manuscript on a similar structure before publication and Lalitha Kodandapani, Robert Campbell, and Susanne Brewerton for discussion. The authors gratefully acknowledge the staff at the LRL-CAT beamline at APS, Stephen Wasserman, John Koss, and Laura Morisco, for data collection. Use of the Advanced Photon Source at Argonne National Laboratory was supported by the U. S. Department of Energy, Office of Science, Office of Basic Energy Sciences , under contract No. DE-AC02-06CH11357. The atomic coordinate file has been deposited RCSB with the PDB code 4MI5. Corresponding author contact: telephone, 858-638-8801; email, luz_john@lilly.com.
PY - 2013/12/19
Y1 - 2013/12/19
N2 - 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.
AB - 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.
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U2 - 10.1371/journal.pone.0084147
DO - 10.1371/journal.pone.0084147
M3 - Article
C2 - 24367637
AN - SCOPUS:84893182116
SN - 1932-6203
VL - 8
JO - PloS one
JF - PloS one
IS - 12
M1 - e84147
ER -