Differential functions of FANCI and FANCD2 ubiquitination stabilize ID2 complex on DNA

Rennie, Martin L.; Lemonidis, Kimon; Arkinson, Connor; Chaugule, Viduth K.; Clarke, Mairi; Streetley, James; Spagnolo, Laura; Walden, Helen

doi:10.1101/2020.02.03.931576

Cited by 12 publications

(67 citation statements)

References 45 publications

(88 reference statements)

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“…Our cell-based assays suggested an interaction between WRNIP1 and the FANCD2/FANCI complex, and using highly purified recombinant proteins, we demonstrated a direct protein-protein interaction with the FANCD2/FANCI complex, as well as monoubiquitinated FANCD2 in complex with FANCI, or Ub-FANCD2/FANCI. Recent work has demonstrated how monoubiquitination of the FANCD2/FANCI complex leads to a substantial conformational change, creating a more stable ring-like structure by which the complex surrounds the DNA ( Alcón et al., 2020 ; Rennie et al., 2020 ; Tan et al., 2020 ; Wang et al., 2020 ). An important part of the conformational change entails rearrangement of the C-terminal Tower domain of FANCD2, which previously was found to be critical for the activity of the FANCD2/FANCI complex via its interaction with DNA ( Liang et al., 2016 ).…”

Section: Discussionmentioning

confidence: 99%

WRNIP1 Is Recruited to DNA Interstrand Crosslinks and Promotes Repair

et al. 2020

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Summary The Fanconi anemia (FA) pathway repairs DNA interstrand crosslinks (ICLs). Many FA proteins are recruited to ICLs in a timely fashion so that coordinated repair can occur. However, the mechanism of this process is poorly understood. Here, we report the purification of a FANCD2-containing protein complex with multiple subunits, including WRNIP1. Using live-cell imaging, we show that WRNIP1 is recruited to ICLs quickly after their appearance, promoting repair. The observed recruitment facilitates subsequent recruitment of the FANCD2/FANCI complex. Depletion of WRNIP1 sensitizes cells to ICL-forming drugs. We find that ubiquitination of WRNIP1 and the activity of its UBZ domain are required to facilitate recruitment of FANCD2/FANCI and promote repair. Altogether, we describe a mechanism by which WRNIP1 is recruited rapidly to ICLs, resulting in chromatin loading of the FANCD2/FANCI complex in an unusual process entailing ubiquitination of WRNIP1 and the activity of its integral UBZ domain.

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Section: Discussionmentioning

confidence: 99%

WRNIP1 Is Recruited to DNA Interstrand Crosslinks and Promotes Repair

et al. 2020

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“…Four concurrent studies also demonstrated a requirement for FANCD2 Ub but not FANCI Ub in DNA clamping, for human, chicken and Xenopus homologs [ 21 , 24 , 25 , 33 ]. The cryo-EM structures of di-monoubiquitinated human FANCI–FANCD2 (FANCI Ub –FANCD2 Ub , PDB: 6VAE , 3.8 Å) reveals why this is the case ( Figure 2 B) [ 25 ].…”

Section: Introductionmentioning

confidence: 99%

“…This new interface created by ubiquitination rationalizes a proposed mechanism by which deubiquitination is controlled. FANCD2 Ub –FANCI is preferentially deubiquitinated compared with FANCD2 Ub –FANCI Ub [ 17 , 33 ], even though the two structures are essentially identical. The only main difference is the occlusion of the USP1 interaction site in FANCD2 [ 37 ] by the presence of the ubiquitin now located on the opposing FANCI [ 25 , 33 ].…”

Section: Introductionmentioning

confidence: 99%

“…FANCD2 Ub –FANCI is preferentially deubiquitinated compared with FANCD2 Ub –FANCI Ub [ 17 , 33 ], even though the two structures are essentially identical. The only main difference is the occlusion of the USP1 interaction site in FANCD2 [ 37 ] by the presence of the ubiquitin now located on the opposing FANCI [ 25 , 33 ]. A patient associated mutation in FANCI (R1285Q) may cause this protective mechanism to break down, as FANCI R1285Q mutant displayed a two-fold faster deubiquitination of FANCD2 [ 25 ].…”

Section: Introductionmentioning

confidence: 99%

“…A patient associated mutation in FANCI (R1285Q) may cause this protective mechanism to break down, as FANCI R1285Q mutant displayed a two-fold faster deubiquitination of FANCD2 [ 25 ]. In cells, unloading of FANCD2 Ub –FANCI Ub may be instigated by DVC1-p97 [ 38 ] but removal of DNA from the FANCD2 Ub –FANCI Ub complex in vitro is sufficient to again permit the activity of USP1:UAF1 on the complex [ 17 , 33 ] highlighting the critical role that DNA plays in stabilizing the structure of the heterodimer. Importantly, the structural changes with-respect to DNA suggest that FANCD2 Ub –FANCI Ub complex will lose its structure-specific binding activity compared with FANCI–FANCD2, which initially has preference for branched DNA molecules [ 25 ].…”

Section: Introductionmentioning

confidence: 99%

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Structural insight into FANCI–FANCD2 monoubiquitination

Tan

Deans

2020

Essays in Biochemistry

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The Fanconi anemia (FA) pathway coordinates a faithful repair mechanism for DNA damage that blocks DNA replication, such as interstrand cross-links. A key step in the FA pathway is the conjugation of ubiquitin on to FANCD2 and FANCI, which is facilitated by a large E3 ubiquitin ligase complex called the FA core complex. Mutations in FANCD2, FANCI or FA core complex components cause the FA bone marrow failure syndrome. Despite the importance of these proteins to DNA repair and human disease, our molecular understanding of the FA pathway has been limited due to a deficit in structural studies. With the recent development in cryo-electron microscopy (EM), significant advances have been made in structural characterization of these proteins in the last 6 months. These structures, combined with new biochemical studies, now provide a more detailed understanding of how FANCD2 and FANCI are monoubiquitinated and how DNA repair may occur. In this review, we summarize these recent advances in the structural and molecular understanding of these key components in the FA pathway, compare the activation steps of FANCD2 and FANCI monoubiquitination and suggest molecular steps that are likely to be involved in regulating its activity.

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DDRugging glioblastoma: understanding and targeting the DNA damage response to improve future therapies

Rominiyi

Collis

2021

Molecular Oncology

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Glioblastoma is the most frequently diagnosed type of primary brain tumour in adults.These aggressive tumours are characterised by inherent treatment resistance and disease progression, contributing to ~190,000 brain-tumour related deaths globally each year. Current therapeutic interventions consist of surgical resection followed by radiotherapy and temozolomide chemotherapy, but average survival is typically around 1 year, with less than 10% of patients surviving more than 5 years. Recently, a fourth treatment modality of intermediate-frequency low-intensity electric fields [called tumourtreating fields ( TTFields)] was clinically approved for glioblastoma in some countries after it was found to increase median overall survival rates by ~5 months in a phase III randomised clinical trial. However, beyond these treatments, attempts to establish more effective therapies have yielded little improvement in survival for patients over the last 50 years. This is in contrast to many other types of cancer and highlights glioblastoma as a recognised tumour of unmet clinical need. Previous work has revealed that glioblastomas contain stem-cell-like subpopulations that exhibit heightened expression of DNA damage

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Differential functions of FANCI and FANCD2 ubiquitination stabilize ID2 complex on DNA

Cited by 12 publications

References 45 publications

WRNIP1 Is Recruited to DNA Interstrand Crosslinks and Promotes Repair

WRNIP1 Is Recruited to DNA Interstrand Crosslinks and Promotes Repair

Structural insight into FANCI–FANCD2 monoubiquitination

DDRugging glioblastoma: understanding and targeting the DNA damage response to improve future therapies

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