Fig. 4: Rpd3S concurrently participates in NCP engagement along with Hho1.

a Top view (left) and side view (right) of the cryo-EM map of Rpd3S co-existing with Hho1 linker histone on the NCP. b Focus refinement of potential Hho1 density with nucleosome revealed clear helical densities that can accommodate the Hho1 globular domain structure. c Particle subtraction of potential Rpd3S density and ab-initial reconstruction revealed an ~ 6 Å map that can unambiguously accommodate the apo Rpd3S complex structure. d EMSA showed Rpd3S co-existing with Hho1 in NCP^187bp. 150 nM NCP^187bp without or with 37.5 μM Hho1 in 7 μL reaction system were shown in lanes 1 to 2 as controls; the Hho1 titration (0 μM, 12.5 μM, 37.5 μM, 112.5 μM, lanes 3 to 6) promoted the bands of Rpd3S–NCP^187bp to shift up, resulting in the formation of Rpd3S–NCP^187bp–Hho1. e Comparison of exit DNA linker angles between the linker tightening state and the Hho1 co-existing state.