This PhD project will address DNA repair mechanisms, specifically the cellular response to DNA crosslinks and replication stress. The Fanconi Anaemia (FA) pathway is at the centre of DNA crosslink repair and involves a large multiprotein E3 ubiquitin ligase (the FA core complex) and the FANCD2-FANCI heterodimer. Deregulation of this pathway can lead to human disease. We aim to understand the molecular mechanisms of crosslink repair at stalled replication forks. Recent work from my group using biochemical reconstitution, single molecule imaging and cryoEM has shown that FANCD2-FANCI forms a DNA clamp that diffuses on double-stranded DNA. By sliding along the DNA, FANCD2-FANCI monitors its integrity and stalls when it reaches a single-stranded gap. This identifies and protects the DNA lesion, and promotes recruitment of downstream repair factors. This project will involve reconstitution of protein-DNA assemblies, identification of protein interactions and determination of structures using cryoEM.
Please contact me for more information and to discuss projects.
Funding Notes
Funding information is available on the institution website.
References
Alcón P, Kaczmarczyk AP, Ray KK, Liolios T, Guilbaud G, Sijacki T, Shen Y, McLaughlin SH, Sale JE, Knipscheer P Rueda DS*, Passmore LA. (2024) FANCD2-FANCI surveys DNA and recognizes double- to single- stranded junctions. Nature 632, 1165-1173.
Sijacki T, Alcón P, Chen ZA, McLaughlin SH, Shakeel S, Rappsilber J, Passmore LA. (2022) The DNA-damage kinase ATR activates the FANCD2-FANCI clamp by priming it for ubiquitination. Nat Struct Mol Biol 29, 881–890
Alcón P†, Shakeel S†, Chen ZA, Rappsilber J, Patel KJ, Passmore LA. (2020) FANCD2–FANCI is a clamp stabilized on DNA by monoubiquitination of FANCD2 during DNA repair. Nat Struct Mol Biol 27, 240–248
Shakeel S†, Rajendra E†, Alcón P, O'Reilly F, Chorev DS, Maslen S, Degliesposti G, Russo CJ, He S, Hill CH, Skehel JM, Scheres SHW, Patel KJ, Rappsilber J, Robinson CV, Passmore LA. (2019) Structure of the Fanconi anemia monoubiquitin ligase complex. Nature 575, 234-237
