CryoEM Structure INO80core Hexasome complex Rvb core refinement state2

Hexasome-INO80 complex reveals structural basis of noncanonical nucleosome remodeling.

Real-space refinement in PHENIX for cryo-EM and crystallography.

New tools for automated cryo-EM single-particle analysis in RELION-4.0.

ISOLDE: a physically realistic environment for model building into low-resolution electron-density maps.

30mM HEPES, pH7.5
50mM NaCl
0.25mM CaCl2
0.25mM DTT
2mM ADP
3.3mM MgCl2
10mM NaF
2mM AlCl3
0.05% octyl-beta-glucoside

11-subunit ct INO80 contains two modules (core and Arp8 module)
Each module was picked and analyzed separately
The core module + hexasome has an overall weight of
0.861MDa
The 11-subunit ct INO80 + hexasome has an overall weight of 1.1MDa

Ino80, Ies2, Ies6, Ies4,Arp6, Rvb1, Rvb2, Arp8, Arp4, Actin, Taf14
Hexasome
DNA, 2xH3, 2xH4, H2A, H2B

INO80 core module in complex with hexasome

Particles were initially picked by WARP to generate an initial model, which was subsequently used for the 3D template picking

11-subunit ctINO80 reconstituted with hexasome

wait time of 5s, blot force at 3, and a blot time of 2s with Whatman blotting paper (Cytiva, CAT No. 10311807)

Chromatin-remodeling complex subunit IES6

EIPOD fellowship under Marie Sklodowska-Curie Actions COFUND

ATP-dependent chromatin remodeler, DNA BINDING PROTEIN