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. Author manuscript; available in PMC: 2019 Feb 15.
Published in final edited form as: Mol Cell. 2018 Feb 15;69(4):677–688.e9. doi: 10.1016/j.molcel.2018.01.028

Figure 4. Deletion of nhp10 results in impaired DNA length sensing by INO80.

Figure 4

(A) Left, denaturing gels showing purifications of Ino80-FLAG(WT), Ino80-FLAG(Δ2-200-ino80) and Ino80-FLAG(Δnhp10). Green and purple stars represent where the Ino80 ATPase and Ies3, a major component of the Nhp10 module run on the gel, respectively. Right, schematic illustrating the compositions of mutant INO80 complexes purified from Δnhp10 and Δ2-200-ino80 strains.

(B) Remodeling rate constants measured by ensemble FRET for 0/40, 0/60 and 0/80 nucleosomes and the various INO80 mutants described in A. These assays were performed under single turnover conditions and with saturating enzyme and ATP.

(C) Native gel remodeling of 0/40 nucleosomes by INO80(Δnhp10) and INO80(Δ2-200-ino80).

These assays were performed under single turnover conditions and with saturating enzyme and ATP. Gel remodeling of 0/60 and 0/80 nucleosomes are shown in Figure S5.

(D) Quantification of fraction cut by REA on 0/80 nucleosomes, with either INO80(WT) or INO80(Δnhp10).

(E) Same as D but with 0/40 nucleosomes. For 0/60 nucleosomes, see Figure S5. Note difference in x-axis from D.

(F) Rate constants measured by REA for 0/40, 0/60 and 0/80 nucleosomes with INO80(WT) or INO80(Δnhp10). These assays were performed under single turnover conditions and with saturating enzyme and ATP. Data in (B) and (F) represent means±S.E.M. for three replicates.