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. Author manuscript; available in PMC: 2022 Nov 16.
Published in final edited form as: Biochemistry. 2021 Nov 1;60(45):3385–3397. doi: 10.1021/acs.biochem.1c00473

Figure 7.

Figure 7.

DNA binding site of Ycs4 resides at the C-terminus. (A) Gel shift assay of WT and mutant Ycs4. (B) Apparent Kd values of Ycs4 mutants. Mutants 1104/08/09/20, 1048/50, 1108/09/20, 1104/08/09, and 1104/20 displayed a decline in DNA binding. (C) Titration of DNA for the stimulation of ATPase rate of the holoenzyme. Lines are spline curves connecting data points. (D) ATP hydrolysis of the holoenzyme condensin in the presence of DNA fit to a two-state binding model (shown with lines), which postulates that DNA binding to the non-SMC subunit activates the Smc2/4 ATPase rate, whereas the non-SMC subunit with mutant Ycs4 is impaired in DNA binding and therefore unable to stimulate the SMC ATPase rate. (E) Correlation between DNA binding, ATP hydrolysis, cell viability, and overproduction toxicity. (F) Sequence alignment of the DNA binding domain of Ycs4 from S. cerevisiae (Sc), Candida auris (Cu), C. albicans (Ca), C. thermophilum (Ct), S. pombe (Sp), Homo sapiens (Hs), and Mus musculus (Mm). Lysines implicated in DNA binding are labeled below the graph.