CryoEM Structure INO80core Hexasome complex composite model state1

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.

This is a composite map composed of focused refinements. We provide a overall reference map, individual cryoEM maps of the focused refinements as well as individually refined structures as separate EMDB and pdb entries.

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

FISHIONE Instruments, Nanoclean
10% O2
90% Argon

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)

N-terminal truncated Ino80 with C-terminal 2xFlag Tag

Chromatin-remodeling complex subunit IES6

Histone H3/a,Histone H3/b,Histone H3/c,Histone H3/d,Histone H3/f,Histone H3/h,Histone H3/i,Histone H3/j,Histone H3/k,Histone H3/l

Histone H2B.1 A,Histone H2B.a,H2B/a,Histone H2B.g,H2B/g,Histone H2B.h,H2B/h,Histone H2B.k,H2B/k,Histone H2B.l,H2B/l

H3C1, H3FA, HIST1H3A, H3C2, H3FL, HIST1H3B, H3C3, H3FC HIST1H3C, H3C4, H3FB, HIST1H3D, H3C6, H3FD, HIST1H3E, H3C7, H3FI, HIST1H3F, H3C8, H3FH, HIST1H3G, H3C10, H3FK, HIST1H3H, H3C11, H3FF, HIST1H3I, H3C12, H3FJ, HIST1H3J

HIST1H4A, H4/A, H4FA, HIST1H4B, H4/I, H4FI, HIST1H4C, H4/G, H4FG, HIST1H4D, H4/B, H4FB, HIST1H4E, H4/J, H4FJ, HIST1H4F, H4/C, H4FC, HIST1H4H, H4/H, H4FH, HIST1H4I, H4/M, H4FM, HIST1H4J, H4/E, H4FE, HIST1H4K, H4/D, H4FD, HIST1H4L, H4/K, H4FK, HIST2H4A, H4/N, H4F2, H4FN, HIST2H4, HIST2H4B, H4/O, H4FO, HIST4H4

HIST1H2BC, H2BFL, HIST1H2BE, H2BFH, HIST1H2BF, H2BFG, HIST1H2BG, H2BFA, HIST1H2BI, H2BFK

EIPOD fellowship under Marie Sklodowska-Curie Actions COFUND
INO80 core bound to hexasome composite model of state 1
Model for focused map / local refinement map
ATP-dependent chromatin remodeler, DNA BINDING PROTEIN