Products/Services Used | Details | Operation |
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Mammalian Expression System> | Full length BARD1 was cloned into pY3H-Ade2 vector by Genscript...Codon optimized SMARCAD1-CUE domains (amino acids 98-318) WT and CUEm (L168E F169E L195E L196E F263E A285E L286E) were custom synthesized by Genscript and sub-cloned into pET15b. ..TGGTTTtcgCCaCGtAGTAAG) and was synthesized by Genscript for WT and L44R variants. ..Human HIST1H2AC was cloned in frame with an N terminal HA tag in pCDNA3.1+ by GenScript...The sequence of all constructs was verified by GenScript....Human SMARCAD1 cDNA was cloned in frame with a N terminal myc tag into pCDNA5/FRT/TO by gene synthesis (GenScript). | Get A Quote |
The opposing activities of 53BP1 and BRCA1 influence pathway choice in DNA double-strand-break repair. How BRCA1 counteracts the inhibitory effect of 53BP1 on DNA resection and homologous recombination is unknown. Here we identify the site of BRCA1-BARD1 required for priming ubiquitin transfer from E2∼ubiquitin and demonstrate that BRCA1-BARD1's ubiquitin ligase activity is required for repositioning 53BP1 on damaged chromatin. We confirm H2A ubiquitination by BRCA1-BARD1 and show that an H2A-ubiquitin fusion protein promotes DNA resection and repair in BARD1-deficient cells. BRCA1-BARD1's function in homologous recombination requires the chromatin remodeler SMARCAD1. SMARCAD1 binding to H2A-ubiquitin and opt... More