Fig. 1: Bre1 interacts with Rad51 in vivo and in vitro.
From: Bre1/RNF20 promotes Rad51-mediated strand exchange and antagonizes the Srs2/FBH1 helicases
a Table showing the Bre1-interacting proteins identified by mass spectrometry analysis. The unique peptide number for each protein is indicated. The score is an indicator of reliability. b, c, h Immunoprecipitation and Western blot analysis of the interaction between Rad51-3xHA and Bre1-3xFLAG or bre1-EK2A-3xFLAG at indicated conditions. GAPDH was used as a loading control. Yeast cells were treated with phleomycin (20 µg/ml) for 2 h or MMS (0.1%) for 1 h before collection. Asynchronized cells were used as control. d, f, g GST pull-down assays showing the interaction between the GST-tagged WT or mutant Bre1 and 6xHis-Rad51. GST was used as a negative control. The bottom panel shows Coomassie blue staining of GST-tagged proteins used for the experiment. The upper panel is the Western blot showing the levels of 6xHis-Rad51. The sizes of proteins are indicated by the molecular-weight markers. e Scheme showing the full-length or truncated Bre1 proteins used for pull-down assays. Dot lines represent deleted regions, while the gray bars represent the truncated Bre1 proteins. The positions for these truncations are indicated. The Rad51-interacting motif of Bre1 is marked by the black bar. Source data are provided as a Source Data file.
