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A table has been added (Fig. 14A) to correlate EM structure and function.
It was not clear whether this funnel is seen in the putative outward facing conformation derived from the 3D EM structure, and the authors do not discuss the significance of its dimensions.
It is, in fact, possible to see clear density for CTP and ADP in the reconstruction; we have clarified this by including in Figure 3 figure supplement 2 a difference map between the EM structure and a calculated model that lacks CTP and ADP.
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EM structures and schematic of the catalytic core sub-complex containing Snf2, Arp7/9 and Rtt102 that are fitted by the crystal structure of Snf2 HSA-Arp7-Arp9 and Rtt102 (PDB ID: 4I6M) (Schubert et al., 2013) and Snf2 post-HSA-ATPase-SnAc domains (PDBID: 5HZR) (Xia et al., 2016) highlighting the substantial mobility of the ATPase domain (bottom panel).
The central cavity is closed at the external face, a finding that was also observed in the EM structures and from cross-linking data for cysteine containing mutants of P-gp.
A recent cryo-EM structure and model of the yeast PIC appears consistent with such accommodation (Murakami et al. 2013).
Are there any differences in the rotamer conformations between your cryo-EM structure and the 1.9 Å X-ray structure?
(B ) Superposition of helices H5, H6, H12 and H13 in the MjNhaP1 EM structure (blue) and corresponding helices in TtNapA (green).
(A ) Sections through protomer volumes of the MjNhaP1 EM structure (left) and the outward-open TtNapA structure (Lee et al., 2013) (right).
The coiled-coil, OB, ATPase domains and the HbYX motif are well defined in crystal and cryo-EM structures, and their primary sequences are highly conserved through evolution (Djuranovic et al., 2009; Chen et al., 2016; Huang et al., 2016; Schweitzer et al., 2016).
In contrast, Δ rimM predominantly contains Group III intermediates in which the head domain is anchored and only partially formed, consistent with head-formation defects observed in cryo-EM structures and by in vivo 16S hydroxyl radical footprinting (Clatterbuck Soper et al., 2013; Guo et al., 2013; Leong et al., 2013).
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