Fig. 1: SIRT1 interacts with and deacetylates SAMHD1 in response to DSBs. | Nature Communications

Fig. 1: SIRT1 interacts with and deacetylates SAMHD1 in response to DSBs.

From: SAMHD1 deacetylation by SIRT1 promotes DNA end resection by facilitating DNA binding at double-strand breaks

Fig. 1: SIRT1 interacts with and deacetylates SAMHD1 in response to DSBs.The alternative text for this image may have been generated using AI.

a Indicated cell lines were treated with or without 5 μM CPT or 10 Gy IR and harvested after 4 h (hrs). Protein lysates were immunoprecipitated (IP’ed) with IgG or anti-SAMHD1 antibody. Input and IP’ed proteins were run on SDS-PAGE gel and western blotted with indicated antibodies. b 293T cells were pre-treated with either 0.5 μM TSA or 10 mM NAM for 1 h and then additionally treated for 4 h in the presence of 10 Gy IR. Protein lysates were IP’ed with anti-SAMHD1 antibody. Input and IP’ed lysates were separated by SDS-PAGE and immunoblotted with indicated antibodies. c, d 293T cells or 293T cells overexpressing SAMHD1-GFP (c) or SIRT1-FLAG (d) were treated with or without 10 Gy IR and harvested after 4 h. Protein lysates processed from the cells were IP’ed with the indicated antibody. Input and IP’ed proteins were run on SDS-PAGE and immunoblotted with indicated antibodies. e 293T protein lysates treated with or without IR were IP’ed with the indicated antibodies, run on SDS-PAGE, and immunoblotted with anti-SIRT1 and SAMHD1 antibodies. f Expression level of the indicated sirtuin proteins in 293T cells transfected with non-targeting siRNA (NT) or siRNA against the indicated sirtuin (SIRT1, T1; SIRT2; T2; SIRT6; T6; SIRT7; T7) was determined by western blot. Three days post-transfection, cells were incubated for 4 h with or without 10 Gy IR and processed for IP with anti-SAMHD1 antibody. Input and pull-down proteins were separated by SDS-PAGE and immunoblotted with indicated antibodies (bottom). g 293T cells were silenced for SIRT1 or a NT control or treated with the SIRT1 inhibitor Ex-527 (1 μM) where indicated for 1 h, treated with or without 10 Gy IR, and harvested 4 h later. Protein lysates were IP’ed with anti-SAMHD1 antibody. Input and IP’ed proteins were run on SDS-PAGE and immunoblotted with indicated antibodies. h 293T cells over-expressing wild-type (WT) or catalytically-dead (HY) SIRT1 were treated with Ex-527 and IR and processed as described in g. Source data are provided as a Source Data file.

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