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Setiaputra Lab
Every cell maintains genome stability
Let’s figure out how.
In the Setiaputra lab, we focus on exploring the molecular mechanisms underlying DNA repair pathway choice. There are multiple potential pathways that respond to DNA damage, and which pathway is brought to bear carries profound implications in the toxicity and mutational outcomes caused by specific genotoxic insults. The molecular basis of the cellular decisions leading to a specific repair trajectory is poorly understood. We use mammalian cell culture combined with biochemistry, cell biology, genomics, and computational biology to address this gap in understanding. Our ultimate goal is to leverage fundamental DNA repair research to identify novel targets and paradigms in targeted cancer therapy and gene editing.
Our current areas of interest are:
- Understanding the mechanism of action of shieldin. BRCA1-mutated breast and ovarian cancers are extremely sensitive to the chemotherapeutic PARP inhibitors. However, loss of the recently discovered shieldin complex results in PARP inhibitor resistance in BRCA1-mutated cells. Though we know that BRCA1 and shieldin promote two competing DNA repair pathways, how shieldin facilitates PARP inhibitor toxicity is not understood. We will tackle this riddle using biochemical reconstitution and cellular biology to understand shieldin’s molecular mechanism of action.
- Monitoring DNA double-strand break repair kinetics in situ. Our understanding of the molecular basis of DNA repair is largely derived from in vitro reconstitution using purified proteins. This often fails to capture the complex regulatory mechanisms influencing DNA repair inside the cell. To get a more accurate picture of DNA repair kinetics, we will use cutting edge time-resolved DNA break sequencing technology to monitor double-strand break repair progression at base-pair resolution as it occurs inside the nucleus.
- Computational analysis of DNA repair networks. DNA repair occurs in extremely crowded compartments where thousands of repair proteins are concentrated. These proteins communicate extensively with each other to influence DNA repair outcomes. To dissect these communication networks, we will use the protein interaction prediction machine learning model AlphaFold-Multimer to mine the DNA repair interactome and discover novel protein binding interfaces that regulate DNA repair.
For more information, please visit our research lab website.
Selected Publications
- An AlphaFold2 map of the 53BP1 pathway identifies a direct SHLD3–RIF1 interaction critical for shieldin activity. Sifri C, Hoeg L, Durocher D, Setiaputra D. EMBO Reports (2023). doi:10.17632/dj2kv8zzxy.
- RIF1 acts in DNA repair through phosphopeptide recognition of 53BP1. Setiaputra D, Escribano-Diaz C, Reinert JK, Sadana P, Zong D, Callen E, Sifri C, Seebacher J, Nussenzweig A, Thomä NH, Durocher D. Molecular Cell (2022). doi:10.1016/j.molcel.2022.01.025.
- Shieldin—the protector of DNA ends. Setiaputra D, Durocher D. EMBO Reports (2019). doi:10.15252/embr.201847560.
- The shieldin complex mediates 53BP1-dependent DNA repair. Noordermeer SM, Adam A*, Setiaputra D*, Barazas M, Pettitt SJ, Ling AK, Olivieri M, Álvarez-Quilón A, Moatti N, Zimmermann M, Annunziato S, Krastev DB, Song F, Brandsma I, Frankum J, Brough R, Sherker A, Landry S, Szilard RK, Munro MM, McEwan A, Goullet de Rugy T, Lin ZY, Hart T, Moffat J, Gingras AC, Martin A, van Attikum H, Jonkers J, Lord CJ, Rottenberg S, Durocher D. Nature (2018). doi:10.1038/s41586-018-0340-7. *Co-first authors.