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- 2019
CTDP1 regulates breast cancer survival and DNA repair through BRCT-specific interactions with FANCIDOI: 10.1038/s41420-019-0185-3 Keywords: Breast cancer, Protein-protein interaction networks Abstract: a Representation of CTDP1 and its modular domain organization. The BRCT region cloned for TAP–MS experiments is indicated between amino acids 619–738. b Workflow diagram of the TAP–MS experiment. c SAINT output for Bayesian false discovery rate (SAINT-BFDR) was used to create a high confidence interaction list. Graph represents the fold change in protein representation between CTDP1 BRCT purification and control and the number of proteins at binned intervals of 0.5 SAINT-BFDR scores. d TAP–MS experiments generate a comprehensive interaction network of 103 proteins interacting with the CTDP1 BRCT domain. Red outlined nodes indicate proteins with GO annotations for DNA damage (GO:0006281) and DNA replication (GO:0006260). Edge color represents source of protein interactions (gray: TAP-MS, blue: BisoGenet). e KEGG pathway enrichment determined by ClueGO of the 103 CTDP1 interacting proteins identified by TAP–MS, excluding CTDP1 itself. The most significant term for each cluster is presented in bold font and term p-value corrected with Bonferroni step down is presented in parentheses. Threshold for visualization was p-value?≤?0.05 and represented inversely proportional to node size. Exact p-values are displayed under most significant group node determined by two-sided hypergeometric test corrected using Bonferroni step down method. f Specificity of protein interactions with the CTDP1 BRCT domain in comparison to 27 other BRCT domain interaction datasets. FANCI and FANCA passing SAINT-BFDR cutoff of ≤0.05 are represented as red circles. g Table detailing central FA pathway proteins’ (FANCA, FANCI, and FANCD2) average spectrum abundance (Ave. Spec.), peptide coverage of protein sequence (Percent Coverage), and SAINT BFDR from TAP-MS. h Analysis of CTDP1 and FANCI expression in the input and immunoprecipation (IP) of control (IgG) or endogenous CTDP1 in complex with FANCI from untreated 293FT cells. i Top: Whole cell lysate (WCL) blot of GFP-FANCI and CTDP1-DDK constructs co-overexpressed in 293FT cells with and without 0.1?μM MMC treated for 24?h. Bottom: IP of DDK-tagged CTDP1 co-immunoprecipitates GFP-tagged FANCI independent of MMC treatmen
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