Figure 4
From: A cell cycle-independent mode of the Rad9-Dpb11 interaction is induced by DNA damage
A Rad9-Dpb11 fusion forces Rad9 recruitment to DSBs and T474 phosphorylation independently of the ‘histone pathway’. (A) The Rad9-Dpb11 fusion blocks resection, also in the absence of Dot1. RPA-ChIP at the indicated positions from an HO-induced DSB (0, 2, 4 and 6 h after HO induction) in WT, dot1Δ, RAD9-DPB11ΔN and RAD9-DPB11ΔN dot1Δ indicates the extent of DNA end resection. FACS profiles in Fig. S4A. (B,C) The Rad9-Dpb11 fusion bypasses the requirement for Dot1, but not for Mec1 and Tel1. Measurement of Rad9-T474 phosphorylation as in Fig. 2A, but in G1-arrested cells expressing the Rad9-Dpb11 fusion in (B) WT and dot1Δ background or (C) WT and mec1Δ tel1Δ background. Immunoblotting against Rad9 or Rad9-T474 phosphorylation. A Pgk1 immunoblot serves as loading control. An asterisk denotes a crossreactive band. FACS profiles in Fig. S4C and E respectively. Strains containing the mec1Δ mutation are in sml1Δ background.
