Smc5/6 Is a Telomere-Associated Complex that Regulates Sir4 Binding and TPE (original) (raw)
Fig 2
Smc5/6 is critical for telomere clustering and Sir4 binding to telomeres.
(A) Rap1-GFP foci in WT (JC1822) and _nse3_-1 (JC3041) cells counted as a measurement for telomere clustering with representative merged images of GFP and DIC channels. (B-C) The number of GFP-Rap1 foci was determined for cells within G1 (unbudded) or S (small budded cells) phases in at least 100 cells for each cell cycle stage, and (D) compared with _mms21_-11 (JC1827) and _smc6_-9 (JC2710). (E-F) Western blot analysis and immunofluorescence staining using α-Myc antibody (green in IF) to detect Sir4Myc in WT (JC3433), _nse3_-1 (JC3452), mms21-11 (JC3597), and smc6-9 (JC2907) cells with DAPI staining shown in blue. (G) ChIP was performed on Sir4Myc as in Fig 1E from asynchronous cultures and in more than one isogenic strain if available. The fold enrichment for each strain is calculated for n≥3 experiments with the mean ± SD at three native subtelomeres (Tel1L, Tel6R and Tel15L). The p values < 0.05 from a two-tailed _t_-test are indicated by (*) for wild type (JC2671 and JC3433), _nse3_-1 (JC3452 and JC3849), _mms21_-11 (JC3597), and _smc6_-9 (JC2907 and JC3087) and non-tagged (nt) control strains included wild type (JC470), _nse3_-1 (JC3607), _mms21_-11 (JC1879), and _smc6_-9 (JC1358).