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. 2016 Feb 18;61(4):563–574. doi: 10.1016/j.molcel.2016.01.026

Figure 7.

Figure 7

Disengagement of NScc1 from Smc3 Requires a Single Round of ATP Hydrolysis

(A) Strains K23070 (MATa SMC3 URA3::SMC3 S1043C-HA6 MYC3-SCC1(TEV268) YEp-PGAL1-TEV), K23067 (MATa SMC3 URA3::smc3 S1043C E1155Q-HA6::URA3 MYC3-SCC1(TEV268) YEp-PGAL1-TEV), and K23068 (MATa SMC3 SMC3 S1043C-HA6 MYC3-SCC1(TEV268) wpl1Δ YEp-PGAL1 TEV) were treated and analyzed as described in Figure 2B.

(B) Strains K24217 (MATa SMC3 URA3::SMC3 S1043C-PK6::URA3 MYC3-SCC1(TEV268)), K24218 (MATa SMC3 URA3::smc3K112 K113Q S1043C-PK6 MYC3-SCC1(TEV268)), K24219 (MATa SMC3 URA3::smc3E1155Q S1043C-PK6 MYC3-SCC1(TEV268)), and K24220 (MATa SMC3 URA3::smc3K112 K113Q E1155Q S1043C-PK6 MYC3-SCC1(TEV268)) were grown and analyzed as described in Figure 6B.

(C) Exponentially growing strains K23070, K23068, K24911 (SMC3 smc3L1126V S1043C::LEU2 MYC3-SCC1(TEV268)), and K24523 (SMC3 S1043C::ADE2 MYC3-SCC1(TEV268) smc1 L1129V) were treated as described in Figure 6B and analyzed by western blotting using anti-MYC antibodies.

(D) Shown is a model for how releasing activity dissociates Scc1-NTD from Smc3’s coiled coil leading to escape of entrapped DNAs in a process involving ATP-dependent engagement of SMC ATPase heads. Acetylation of Smc3 residues K112 and K113 is suggested to inhibit ATP-dependent head engagement.