Skip to main content
. 2017 Feb 1;45(7):3844–3859. doi: 10.1093/nar/gkx065

Figure 2.

Figure 2.

The RQC domain of WRN responds to oxidative damage at telomeres but not at genome sites. (A) Schematic representation of the DDR of a full length (FL) GFP-tagged WRN and truncated or mutated WRN at telomeres and genome sites. (+) means a positive damage response and (−) means no damage response. (B) Three min after 559 nm laser light activation of KR, recruitment of GFP-tagged truncated or mutated WRN proteins to DsRed or KR-TRF1-induced damage at telomeres is shown in U2OS cells. The yellow rectangle indicates an enlarged area showing the colocalization of WRN at the sites of KR-TRF1. (C) Upper panel: recruitment of RQC domain of WRN at telomeres after KR-TRF1 activation in HeLa cells. Lower panel: Scheme of Damage Targeted at Genomic sites method to induce ROS damage specifically at one genome locus of a chromosome is shown. The tetracycline repressor (tetR) fused to KR (tetR-KR) binds a tetracycline response element (TRE) cassette in the defined genome site in U2OS cells. (D) Recruitment of GFP-tagged WRN FL and the RQC domain to 405 nm laser induced damage before (left) and 3 min after (right) 405 nm laser irradiation for 500 ms. The DDR of WRN FL but not the RQC domain to tetR-KR (D) or 405 nm laser (E) is shown with yellow arrowheads. (E) Quantification of accumulation kinetics of GFP-tagged WRN FL and the RQC domain at sites of telomeric damage by the fold increase of the relative intensity after 559 nm laser light irradiation. (F) Dissociation kinetics of GFP-tagged WRN FL and the RQC domain at sites of telomeric damage at 10 min, 24 and 48 h recovery time after 20 min cool white fluorescent bulb light activation of KR. The percentages of co-localization of WRN FL or the RQC domain with KR-TRF1 are shown in the graph. A total of 150 cells in total were counted in three independent experiments; data are represented as mean ± SEM.