TY - JOUR
T1 - A bidentate Polycomb Repressive-Deubiquitinase complex is required for efficient activity on nucleosomes
AU - Foglizzo, Martina
AU - Middleton, Adam J.
AU - Burgess, Abigail E.
AU - Crowther, Jennifer M.
AU - Dobson, Renwick C.J.
AU - Murphy, James M.
AU - Day, Catherine L.
AU - Mace, Peter D.
N1 - Funding Information:
We thank Sam Jamieson for technical assistance with various aspects of the project, Adam Graham for preliminary construct testing, Alan Riboldi-Tunnicliffe for diffraction screening at the Australian Synchrotron, and Michael Algie for helpful discussions. P.D.M. was supported by a Rutherford Discovery Fellowship from the New Zealand government administered by the Royal Society of New Zealand. Support was provided by a Project Grant from the Health Research Council of New Zealand, the Biochemistry Department at the University of Otago, a University of Otago Research Grant (to P.D.M.), the Victorian State Government Operational Infrastructure Support, NHMRC IRIISS grant (9000433), and an NHMRC fellowship (1105754; to J.M.M.).
Publisher Copyright:
© 2018, The Author(s).
PY - 2018/12/1
Y1 - 2018/12/1
N2 - Attachment of ubiquitin to lysine 119 of Histone 2A (H2AK119Ub) is an epigenetic mark characteristic of repressed developmental genes, which is removed by the Polycomb Repressive-Deubiquitinase (PR-DUB) complex. Here we report the crystal structure of the Drosophila PR-DUB, revealing that the deubiquitinase Calypso and its activating partner ASX form a 2:2 complex. The bidentate Calypso–ASX complex is generated by dimerisation of two activated Calypso proteins through their coiled-coil regions. Disrupting the Calypso dimer interface does not affect inherent catalytic activity, but inhibits removal of H2AK119Ub as a consequence of impaired recruitment to nucleosomes. Mutating the equivalent surface on the human counterpart, BAP1, also compromises activity on nucleosomes. Together, this suggests that high local concentrations drive assembly of bidentate PR-DUB complexes on chromatin—providing a mechanistic basis for enhanced PR-DUB activity at specific genomic foci, and the impact of distinct classes of PR-DUB mutations in tumorigenesis.
AB - Attachment of ubiquitin to lysine 119 of Histone 2A (H2AK119Ub) is an epigenetic mark characteristic of repressed developmental genes, which is removed by the Polycomb Repressive-Deubiquitinase (PR-DUB) complex. Here we report the crystal structure of the Drosophila PR-DUB, revealing that the deubiquitinase Calypso and its activating partner ASX form a 2:2 complex. The bidentate Calypso–ASX complex is generated by dimerisation of two activated Calypso proteins through their coiled-coil regions. Disrupting the Calypso dimer interface does not affect inherent catalytic activity, but inhibits removal of H2AK119Ub as a consequence of impaired recruitment to nucleosomes. Mutating the equivalent surface on the human counterpart, BAP1, also compromises activity on nucleosomes. Together, this suggests that high local concentrations drive assembly of bidentate PR-DUB complexes on chromatin—providing a mechanistic basis for enhanced PR-DUB activity at specific genomic foci, and the impact of distinct classes of PR-DUB mutations in tumorigenesis.
UR - https://www.scopus.com/pages/publications/85054077829
U2 - 10.1038/s41467-018-06186-1
DO - 10.1038/s41467-018-06186-1
M3 - Article
C2 - 30258054
AN - SCOPUS:85054077829
SN - 2041-1723
VL - 9
JO - Nature Communications
JF - Nature Communications
IS - 1
M1 - 3932
ER -