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. 2020 Apr 27;9:e55667. doi: 10.7554/eLife.55667

Figure 4. Kin28 phosphorylation of RNA polymerase II CTD is critical for H2A.Z eviction.

(A) Schematic representation shows the three Pol II kinases Kin28, Bur1 and Ctk1 recruited at initiation, early-elongation and elongation phases respectively of Pol II and corresponding phosphorylation of indicated Rpb1 CTD sites. Set1 is the first of the three RNA capping enzymes; it removes γ-phosphate from the RNA 5’end to generate 5’ diphosphate. (B, C, D, E) Normalized histograms and two-component Gaussian fits for H2A.Z-Halo imaged in cells co-depleted for Swc5 along with Kin28 (B), Bur1 (C), Ctk1 (D) and Cet1 (E). (F) Spot-On results show Kin28 is required to evict H2A.Z. All molecules tracked with JF552 dye.

Figure 4.

Figure 4—figure supplement 1. H2A.Z diffusion histograms in cells after single depletion of Kin28.

Figure 4—figure supplement 1.

(A, B) H2A.Z-Halo distributions in Kin28-AA strain without (A) and with (B) depletion of Kin28. (C) Spot-On results for chromatin bound H2A.Z upon Kin28 depletion. (D) Reproduction of Gaussian fits for H2A.Z-Halo distributions in rapamycin-treated Swc5-AA cells (from Figure 2C). (E) H2A.Z-Halo distributions in cells co-depleted for Swc5 and Rrp6. (F) Spot-On results for chromatin-bound H2A.Z upon double depletion of Swc5 and Rrp6. All molecules tracked with JF552 dye.