Investigating the role of e3 ubiquitin ligases in the cancer responses to ionising radiation (ir)
The Ubiquitin Proteasome System (UPS) is a crucial regulator of cell survival in normal conditions and after DNA damage. The function of UPS is dysregulated in cancer, thus providing a large repertoire of targets to exploit for better cancer treatment. While there are more than 1000 genes composing the UPS, the function and mechanisms of action of the majority of them are unknown.
The mission of the D’Angiolella laboratory is to decipher the role of the E3 ubiquitin ligases (Fig.1A), components of the UPS, in cancer pathogenesis and response to treatment with Ionising Radiation . The laboratory collaborates closely with structural biologists to decipher the role of Cul3 ubiquitin ligases (Fig.1B). The laboratory has conducted UPS focused CRISPR screens to have a system-wide view of the role of individual components of the UPS in the cellular responses to IR and cancer pathogenesis.
Investigation into the role of E3s will provide insights in the pathogenetic mechanisms of cancers and pose the basis for drug development through PROteolysis TArgeting Chimeras (PROTACs) and immunomodulatory IMiDs, such as lenalidomide (Fig.1C).
Figure 1. The modular architectures of Cullin-RING E3 Ligases (CRLs). (A) CRL1 ubiquitin ligase complexes are perhaps the best known examples and bind their substrates via an F-box containing subunit. D'Angiolella and his laboratory discovered functions of cyclin F (FBXO1), the founding member of the F-box family of Cullin-RING ubiquitin ligases. (B) BTB-Kelch proteins assemble into CRL3 ubiquitin ligase complexes in which the BTB domain binds to Cullin3 and the Kelch domain forms the substrate recognition domain. (C) Some Cullin-RING ubiquitin ligase subunits, such as Cereblon are bound by “molecular glue” drugs (e.g. lenalidomide) that enable the recruitment of neo-substrates (e.g. Ikaros) for ubiquitylation. Similarly, bifunctional small molecule PROTAC drugs can bind to the E3 ligase VHL to recruit neo-substrates (e.g. oncoproteins) for ubiquitylation and degradation.
Students will be trained in techniques such as high resolution CRISPR screens, biochemical assays and super resolution microscopy. There will be comprehensive training on methods to investigate the ubiquitin system including novel techniques and approaches for PROTACs generation. The laboratory offers the possibility to attend international courses (EMBO and others). Attendance to at least one international conference is guaranteed.
- Chen Z, Wasney GA, Picaud S, Filippakopoulos P, Vedadi M, D'Angiolella V, Bullock AN. Identification of a PGXPP degron motif in dishevelled and structural basis for its binding to the E3 ligase KLHL12. Open Biol. 2020 Jun;10(6):200041. doi: 10.1098/rsob.200041. Epub 2020 Jun 24. PMID: 32574548; PMCID: PMC7333892.
- Burdova K, Yang H, Faedda R, Hume S, Chauhan J, Ebner D, Kessler BM, Vendrell I, Drewry DH, Wells CI, Hatch SB, Dianov GL, Buffa FM, D'Angiolella V. E2F1 proteolysis via SCF-cyclin F underlies synthetic lethality between cyclin F loss and Chk1 inhibition. EMBO J. 2019 Oct 15;38(20):e101443. doi: 10.15252/embj.2018101443. Epub 2019 Aug 19. PMID: 31424118; PMCID: PMC6792013.
- D'Angiolella V, Donato V, Forrester FM, Jeong YT, Pellacani C, Kudo Y, Saraf A, Florens L, Washburn MP, Pagano M. Cyclin F-mediated degradation of ribonucleotide reductase M2 controls genome integrity and DNA repair. Cell. 2012 May 25;149(5):1023-34. doi: 10.1016/j.cell.2012.03.043. PMID: 22632967; PMCID: PMC3616325.