Journal: Nucleic Acids Research

Article Title: Tetrameric INTS6-SOSS1 complex facilitates DNA:RNA hybrid autoregulation at double-strand breaks

doi: 10.1093/nar/gkae937

Figure Lengend Snippet: INTS6 is required for efficient DNA damage repair. ( A ) Left: Representative images of the clonogenic assay in control and INTS6 knockdown cells. The cells were stained and counted after 10 days of growing. Right: Quantification of left. * P ≤ 0.05, ** P ≤ 0.01. ( B ) MTT assay to show cell viability (%) at indicated time points in siCtrl and siINTS6 cells with or without 2 Gy IR treatment. Error bar = mean ± SD, significance was determined using unpaired Student’s t -test, * P ≤ 0.05. ( C ) Left: Drawing of DR-GFP HR reporter strategy. Right: Bar chart shows the efficiency of HR repair in DR-GFP HeLa reporter cells, as measured by FACS. BRCA1 knockdown was used as the positive control. ****P ≤ 0.0001, ***P ≤ 0.001. ( D ) Top: Representative images of comet assay in siCtrl, siINTS6, siRAD51 and si53BP1 cells. siRAD51 and si53BP1 cells were used as positive controls. IR = 5 Gy. Error bar = 100 μm. Bottom: Quantification of top. **** P ≤ 0.0001, ** P ≤ 0.01. ( E ) Model: INTS6 as part of tetrameric SOSS1 complex binds to DNA:RNA hybrids at DSBs and recruits PP2A to dephosphorylate RNAPII. Depletion of INTS6 results in increased levels of DARTs and DNA:RNA hybrids. INTS6 interacts with SETX and is required for its recruitment to damaged sites. SETX, in turn, resolves DNA:RNA hybrids at DSBs facilitating their INTS6-dependent autoregulation. Image was created with Biorender.

Article Snippet: HeLa wild-type (WT) cells were obtained from ATCC.

Techniques: Clonogenic Assay, Control, Knockdown, Staining, MTT Assay, Positive Control, Single Cell Gel Electrophoresis