Journal
EMBO JOURNAL
Volume 40, Issue 22, Pages -Publisher
WILEY
DOI: 10.15252/embj.2019103787
Keywords
E3 ligase; homologous recombination; post-translational modification; replication protein A; small ubiquitin-like modifier
Categories
Funding
- NIH National Institute of General Medical Sciences (NIGMS) [R35 GM118080]
- NIH National Cancer Institute-Cancer Center Support Grant [P30 CA008748]
- Fonds de Recherche du Quebec-Sante
- NIH NIGMS [P41 GM103403, S10 RR029205]
- DOE Office of Science by Argonne National Laboratory [DE-AC02-06CH11357]
- Howard Hughes Medical Institute
Ask authors/readers for more resources
The repair of DNA double-stranded breaks by homologous recombination is dependent on DNA end resection and post-translational modification of repair factors. The SAP domain of the E3 ligase Siz2 plays a key role in promoting SUMO modification of RPA during DNA repair, as highlighted by enzymatic assays and crystal structure analysis. Our results provide insights on how DNA architecture scaffolds a substrate and E3 ligase to promote SUMO modification in the context of DNA repair.
Repair of DNA double-stranded breaks by homologous recombination (HR) is dependent on DNA end resection and on post-translational modification of repair factors. In budding yeast, single-stranded DNA is coated by replication protein A (RPA) following DNA end resection, and DNA-RPA complexes are then SUMO-modified by the E3 ligase Siz2 to promote repair. Here, we show using enzymatic assays that DNA duplexes containing 3' single-stranded DNA overhangs increase the rate of RPA SUMO modification by Siz2. The SAP domain of Siz2 binds DNA duplexes and makes a key contribution to this process as highlighted by models and a crystal structure of Siz2 and by assays performed using protein mutants. Enzymatic assays performed using DNA that can accommodate multiple RPA proteins suggest a model in which the SUMO-RPA signal is amplified by successive rounds of Siz2-dependent SUMO modification of RPA and dissociation of SUMO-RPA at the junction between single- and double-stranded DNA. Our results provide insights on how DNA architecture scaffolds a substrate and E3 ligase to promote SUMO modification in the context of DNA repair.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available