Fig. 3: The UBX domains of IARS1 interact with the RING and BRCT domains of BRCA1 for protein stability.
From: Regulation of BRCA1 stability through the tandem UBX domains of isoleucyl-tRNA synthetase 1
a Schematic illustration of the hypothetical model by which IARS1 UBX domains protect partner proteins from proteasomal degradation. b The candidate proteins that interact with IARS1 were classified into several biological processes using the BioGrid database. c Immunoprecipitation of endogenous BRCA1 was performed with an anti-BRCA1 antibody in HCT116 cells. Co-precipitated endogenous IARS1 was detected by immunoblotting with an anti-IARS1 antibody. d In vitro affinity pull-down analysis of IARS1 with the BRCA1-BARD1 complex via the Flag tag of BRCA1. The supernatant (S), wash (W), and eluate (E) fractions were analyzed by immunoblotting with anti-Flag and anti-His antibodies. e Immunoassay of co-expressed Myc-BRCA1 and Flag-tagged full-length IARS1 or its various domains in HEK293T cells. Flag-IARS1 was immunoprecipitated with an anti-Flag antibody, and co-precipitated Myc-BRCA1 was detected by immunoblotting with an anti-Myc antibody. f In vitro pull-down assay of IARS1 and the following fragments of the BRCA1-BARD1 complex: RING-RING domains, BRCT domain of BRCA1, and BRCT domain of BARD1. g HEK293T cells were co-transfected with wild-type Flag-tagged BRCA1 or deletion mutants (ΔRING or ΔBRCT domain) and HA-tagged IARS1, and then immunoprecipitation was performed with an anti-Flag antibody. Co-precipitated IARS1 was detected by immunoblotting with an anti-HA antibody. h Co-IP analysis for the interaction between endogenous IARS1 and BRCA1 using a BRCA1-specific antibody in nuclear extract. c–h The data are representative of three independent experiments. Source data are provided as a Source Data file.
