Fig. 2
From: Rif1 S-acylation mediates DNA double-strand break repair at the inner nuclear membrane
C466 and C473 are required for Rif1-mediated NHEJ. a Top, Budding yeast Rif1 (1916 amino acid residues) contains four identifiable functional domains: RVxF/SILK PP1-interacting domain; RBM and CTD Rap1-binding motifs; HOOK DNA-binding domain. The Rif1 N-terminal domain (Rif1NTD, residues 1–1322) supports the protein’s function in NHEJ. The positions of cysteine residues, representing potential Rif1 S-acylation sites, are indicated. Bottom, surface representation of Rif1 (residues 177–1283) structure bound with DNA3, identifying surface-exposed cysteines. For mutational analyses, cysteines were grouped by proximity into clusters 1 and 2. C1292 and C71, for which structural data is not available, were included in cluster 1 and 2, respectively. b NHEJ efficiency for Rif1NTD cysteine clusters 1 and 2 mutants, determined as in Fig. 1b after 2 h of HO-endonuclease induction. Data are presented as mean values ± s.e.m. (n = 3 independent experiments). c Viability of the indicated strains in the presence of Zeocin (70 μg/ml). Data are presented as mean values ± s.e.m. (n = 6 independent experiments). Statistical analysis was performed by one-way Anova and a post-hoc Tukey–Kramer multiple comparison test, comparing wild-type to the indicated mutants. PE, plating efficiency; PET0, plating efficiency without HO-endonuclease induction; PERIF1, plating efficiency of RIF1 wild-type reference strain. See also Supplementary Fig. 2. Source data are provided as a Source Data file
