scholarly journals Structure of the Fanconi Anemia Core–UBE2T complex poised to ubiquitinate bound FANCI–FANCD2

2019 ◽  
Author(s):  
Shengliu Wang ◽  
Renjing Wang ◽  
Christopher Peralta ◽  
Ayat Yaseen ◽  
Nikola P. Pavletich
Keyword(s):  
Ubiquitin Ligase ◽  
Fanconi Anemia ◽  
Replication Fork ◽  
Single Copy ◽  
Core Complex ◽  
Interstrand Crosslinks ◽  
Ubiquitination Site ◽  
Pathway Activation ◽  
Dna Interstrand Crosslinks ◽  
Ubiquitin Conjugating Enzyme

ABSTRACTThe Fanconi Anemia (FA) pathway is essential for the repair of DNA interstrand crosslinks (ICLs). The pathway is activated when a replication fork stalls because of an ICL or other replication stress. A central event in pathway activation is the mono-ubiquitination of the FANCI-FANCD2 (ID) complex by the FA Core complex, a ubiquitin ligase of nine subunits. Here we describe the cryo-EM structures of the 1.1 MDa FA Core at 3.1 angstroms, except for the FANCA subunit at 3.4, and of the complex containing Core, ID and the UBE2T ubiquitin conjugating enzyme at 4.2 angstroms. The Core has unusual stoichiometry with two copies of FANCB, FAAP100, FANCA, FAAP20, FANCG, FANCL, but only a single copy of FANCC, FANCE and FANCF. This is due to homodimers of FANCA and FANCB having incompatible 2-fold symmetry, resulting in an overall asymmetric assembly of the other subunits. The asymmetry is crucial, as it prevents the binding of a second FANC-C-E-F sub-complex that inhibits UBE2T recruitment by FANCL, and instead creates an ID binding site. The single active FANCL-UBE2T binds next to the FANCD2 ubiquitination site, prying open the FANCI-FANCD2 interface within which the ubiquitination sites are buried. These structures and biochemical data indicate a single active site ubiquitinates FANCD2 and FANCI sequentially, shedding light on a central event in the FA pathway.

scholarly journals Towards a Molecular Understanding of the Fanconi Anemia Core Complex

Anemia ◽  
2012 ◽  
Vol 2012 ◽  
pp. 1-10 ◽  
Author(s):  
Charlotte Hodson ◽  
Helen Walden
Keyword(s):  
Fanconi Anemia ◽  
Molecular Mechanisms ◽  
Genome Instability ◽  
Genetic Disorder ◽  
Single Gene ◽  
Main Function ◽  
Core Complex ◽  
Interstrand Crosslinks ◽  
Dna Interstrand Crosslinks ◽  
Interstrand Crosslinking

Fanconi Anemia (FA) is a genetic disorder characterized by the inability of patient cells to repair DNA damage caused by interstrand crosslinking agents. There are currently 14 verified FA genes, where mutation of any single gene prevents repair of DNA interstrand crosslinks (ICLs). The accumulation of ICL damage results in genome instability and patients having a high predisposition to cancers. The key event of the FA pathway is dependent on an eight-protein core complex (CC), required for the monoubiquitination of each member of the FANCD2-FANCI complex. Interestingly, the majority of patient mutations reside in the CC. The molecular mechanisms underlying the requirement for such a large complex to carry out a monoubiquitination event remain a mystery. This paper documents the extensive efforts of researchers so far to understand the molecular roles of the CC proteins with regard to its main function in the FA pathway, the monoubiquitination of FANCD2 and FANCI.

scholarly journals Regulation of the Fanconi Anemia DNA Repair Pathway by Phosphorylation and Monoubiquitination

Genes ◽  
2021 ◽  
Vol 12 (11) ◽  
pp. 1763
Author(s):  
Masamichi Ishiai
Keyword(s):  
Dna Repair ◽  
Fanconi Anemia ◽  
Genome Stability ◽  
Short Review ◽  
Repair Pathway ◽  
Binding Partner ◽  
Interstrand Crosslinks ◽  
Pathway Activation ◽  
Dna Interstrand Crosslinks

The Fanconi anemia (FA) DNA repair pathway coordinates a faithful repair mechanism for stalled DNA replication forks caused by factors such as DNA interstrand crosslinks (ICLs) or replication stress. An important role of FA pathway activation is initiated by monoubiquitination of FANCD2 and its binding partner of FANCI, which is regulated by the ATM-related kinase, ATR. Therefore, regulation of the FA pathway is a good example of the contribution of ATR to genome stability. In this short review, we summarize the knowledge accumulated over the years regarding how the FA pathway is activated via phosphorylation and monoubiquitination.

scholarly journals UBE2T, the Fanconi Anemia Core Complex, and FANCD2 Are Recruited Independently to Chromatin: a Basis for the Regulation of FANCD2 Monoubiquitination

2007 ◽  
Vol 27 (24) ◽  
pp. 8421-8430 ◽  
Author(s):  
Arno Alpi ◽  
Frederic Langevin ◽  
Georgina Mosedale ◽  
Yuichi J. Machida ◽  
Anindya Dutta ◽  
...  
Keyword(s):  
Dna Damage ◽  
Ubiquitin Ligase ◽  
Fanconi Anemia ◽  
E3 Ubiquitin Ligase ◽  
S Phase ◽  
Multiprotein Complex ◽  
Core Complex ◽  
Nuclear Core ◽  
Ubiquitin Conjugating Enzyme ◽  
Fancd2 Monoubiquitination

ABSTRACT The Fanconi anemia (FA) nuclear core complex and the E2 ubiquitin-conjugating enzyme UBE2T are required for the S phase and DNA damage-restricted monoubiquitination of FANCD2. This constitutes a key step in the FA tumor suppressor pathway, and much attention has been focused on the regulation at this point. Here, we address the importance of the assembly of the FA core complex and the subcellular localization of UBE2T in the regulation of FANCD2 monoubiquitination. We establish three points. First, the stable assembly of the FA core complex can be dissociated of its ability to function as an E3 ubiquitin ligase. Second, the actual E3 ligase activity is not determined by the assembly of the FA core complex but rather by its DNA damage-induced localization to chromatin. Finally, UBE2T and FANCD2 access this subcellular fraction independently of the FA core complex. FANCD2 monoubiquitination is therefore not regulated by multiprotein complex assembly but by the formation of an active E2/E3 holoenzyme on chromatin.

scholarly journals trans-Fatty acids promote p53-dependent apoptosis triggered by cisplatin-induced DNA interstrand crosslinks via the Nox-RIP1-ASK1-MAPK pathway

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Yusuke Hirata ◽  
Miki Takahashi ◽  
Yuto Yamada ◽  
Ryosuke Matsui ◽  
Aya Inoue ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Mapk Pathway ◽  
Regulatory Protein ◽  
Strand Break ◽  
Mitogen Activated Protein Kinase ◽  
Ros Generation ◽  
Apoptotic Pathway ◽  
Interstrand Crosslinks ◽  
Pathway Activation ◽  
Dna Interstrand Crosslinks

Abstracttrans-Fatty acids (TFAs) are food-derived fatty acids associated with various diseases including cardiovascular diseases. However, the underlying etiology is poorly understood. Here, we show a pro-apoptotic mechanism of TFAs such as elaidic acid (EA), in response to DNA interstrand crosslinks (ICLs) induced by cisplatin (CDDP). We previously reported that TFAs promote apoptosis induced by doxorubicin (Dox), a double strand break (DSB)-inducing agent, via a non-canonical apoptotic pathway independent of tumor suppressor p53 and apoptosis signal-regulating kinase (ASK1), a reactive oxygen species (ROS)-responsive kinase. However, here we found that in the case of CDDP-induced apoptosis, EA-mediated pro-apoptotic action was reversed by knockout of either p53 or ASK1, despite no increase in p53 apoptotic activity. Upon CDDP treatment, EA predominantly enhanced ROS generation, ASK1-p38/c-Jun N-terminal kinase (JNK) mitogen-activated protein kinase (MAPK) pathway activation, and ultimately cell death, all of which were suppressed either by co-treatment of the NADPH oxidase (Nox) inhibitor Apocynin, or by knocking out its regulatory protein, receptor-interacting protein 1 (RIP1). These results demonstrate that in response to CDDP ICLs, TFAs promote p53-dependent apoptosis through the enhancement of the Nox-RIP1-ASK1-MAPK pathway activation, providing insight into the diverse pathogenetic mechanisms of TFAs according to the types of DNA damage.

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

2020 ◽  
Author(s):  
Martin L. Rennie ◽  
Kimon Lemonidis ◽  
Connor Arkinson ◽  
Viduth K. Chaugule ◽  
Mairi Clarke ◽  
...  
Keyword(s):  
Fanconi Anemia ◽  
Large Scale ◽  
The Other ◽  
Molecular Function ◽  
Post Translational Modifications ◽  
Hydrophobic Residues ◽  
Interstrand Crosslinks ◽  
Double Stranded Dna ◽  
Dna Interstrand Crosslinks ◽  
Lysine Residues

AbstractThe Fanconi Anemia (FA) pathway is a dedicated pathway for the repair of DNA interstrand crosslinks, and which is additionally activated in response to other forms of replication stress. A key step in the activation of the FA pathway is the monoubiquitination of each of the two subunits (FANCI and FANCD2) of the ID2 complex on specific lysine residues. However, the molecular function of these modifications has been unknown for nearly two decades. Here we find that ubiquitination of FANCD2 acts to increase ID2’s affinity for double stranded DNA via promoting/stabilizing a large-scale conformational change in the complex, resulting in a secondary “Arm” ID2 interphase encircling DNA. Ubiquitination of FANCI, on the other hand, largely protects the ubiquitin on FANCD2 from USP1/UAF deubiquitination, with key hydrophobic residues of FANCI’s ubiquitin being important for this protection. In effect, both of these post-translational modifications function to stabilise a conformation in which the ID2 complex encircles DNA.

scholarly journals Acetylation modulates the Fanconi anemia pathway by protecting FAAP20 from ubiquitin-mediated proteasomal degradation

2020 ◽  
Vol 295 (40) ◽  
pp. 13887-13901
Author(s):  
Bhavika Nagareddy ◽  
Arafat Khan ◽  
Hyungjin Kim
Keyword(s):  
Fanconi Anemia ◽  
Posttranslational Modifications ◽  
Genome Stability ◽  
Chromosome Instability ◽  
Lysine Residue ◽  
Core Complex ◽  
Ubiquitin E3 Ligase ◽  
Pathway Activation ◽  
Anemia Group ◽  
Inherited Mutations

Fanconi anemia (FA) is a chromosome instability syndrome of children caused by inherited mutations in one of FA genes, which together constitute a DNA interstrand cross-link (ICL) repair, or the FA pathway. Monoubiquitination of Fanconi anemia group D2 protein (FANCD2) by the multisubunit ubiquitin E3 ligase, the FA core complex, is an obligate step in activation of the FA pathway, and its activity needs to be tightly regulated. FAAP20 is a key structural component of the FA core complex, and regulated proteolysis of FAAP20 mediated by prolyl cis-trans isomerization and phosphorylation at a consensus phosphodegron motif is essential for preserving the integrity of the FA core complex, and thus FANCD2 monoubiquitination. However, how ubiquitin-dependent FAAP20 degradation is modulated to fine-tune FA pathway activation remains largely un-known. Here, we present evidence that FAAP20 is acetylated by the acetyltransferase p300/CBP on lysine 152, the key residue that when polyubiquitinated results in the degradation of FAAP20. Acetylation or mutation of the lysine residue stabilizes FAAP20 by preventing its ubiquitination, thereby protecting it from proteasome-dependent FAAP20 degradation. Consequently, disruption of the FAAP20 acetylation pathway impairs FANCD2 activation. Together, our study reveals a competition mechanism between ubiquitination and acetylation of a common lysine residue that controls FAAP20 stability and highlights a complex balancing between different posttranslational modifications as a way to refine the FA pathway signaling required for DNA ICL repair and genome stability.

scholarly journals UHRF1 Is a Sensor for DNA Interstrand Crosslinks and Recruits FANCD2 to Initiate the Fanconi Anemia Pathway

Cell Reports ◽  
2015 ◽  
Vol 10 (12) ◽  
pp. 1947-1956 ◽  
Author(s):  
Chih-Chao Liang ◽  
Bao Zhan ◽  
Yasunaga Yoshikawa ◽  
Wilhelm Haas ◽  
Steven P. Gygi ◽  
...  
Keyword(s):  
Fanconi Anemia ◽  
Fanconi Anemia Pathway ◽  
Interstrand Crosslinks ◽  
Dna Interstrand Crosslinks

scholarly journals FANCD2, FANCJ and BRCA2 cooperate to promote replication fork recovery independently of the Fanconi Anemia core complex

Cell Cycle ◽  
2015 ◽  
Vol 14 (3) ◽  
pp. 342-353 ◽  
Author(s):  
Maya Raghunandan ◽  
Indrajit Chaudhury ◽  
Stephanie L. Kelich ◽  
Helmut Hanenberg ◽  
Alexandra Sobeck
Keyword(s):  
Fanconi Anemia ◽  
Replication Fork ◽  
Core Complex

scholarly journals DNA clamp function of the mono-ubiquitinated Fanconi Anemia FANCI-FANCD2 complex

2019 ◽  
Author(s):  
Renjing Wang ◽  
Shengliu Wang ◽  
Ankita Dhar ◽  
Christopher Peralta ◽  
Nikola P. Pavletich
Keyword(s):  
Fanconi Anemia ◽  
Translesion Synthesis ◽  
The Other ◽  
Replication Forks ◽  
Dna Structures ◽  
Cancer Predisposition Syndrome ◽  
Interstrand Crosslinks ◽  
Branched Dna ◽  
Closed Ring ◽  
Dna Interstrand Crosslinks

ABSTRACTThe FANCI-FANCD2 (ID) complex, mutated in the Fanconi Anemia (FA) cancer predisposition syndrome, is required for the repair of replication forks stalled at DNA interstrand crosslinks (ICL) and related lesions1. The FA pathway is activated when two replication forks converge onto an ICL2, triggering the mono-ubiquitination of the ID complex. ID mono-ubiquitination is essential for ICL repair by excision, translesion synthesis and homologous recombination, but its function was hitherto unknown1,3. Here, the 3.48 Å cryo-EM structure of mono-ubiquitinated ID (IDUb) bound to DNA reveals that it forms a closed ring that encircles the DNA. Compared to the cryo-EM structure of the non-ubiquitinated ID complex bound to ICL DNA, described here as well, mono-ubiquitination triggers a complete re-arrangement of the open, trough-like ID structure through the ubiquitin of one protomer binding to the other protomer in a reciprocal fashion. The structures, in conjunction with biochemical data, indicate the mono-ubiquitinated ID complex looses its preference for ICL and related branched DNA structures, becoming a sliding DNA clamp that can coordinate the subsequent repair reactions. Our findings also reveal how mono-ubiquitination in general can induce an alternate structure with a new function.

scholarly journals Cooperation of the NEIL3 and Fanconi anemia/BRCA pathways in interstrand crosslink repair

2020 ◽  
Vol 48 (6) ◽  
pp. 3014-3028 ◽  
Author(s):  
Niu Li ◽  
Jian Wang ◽  
Susan S Wallace ◽  
Jing Chen ◽  
Jia Zhou ◽  
...  
Keyword(s):  
Fanconi Anemia ◽  
Excision Repair ◽  
Human Cells ◽  
Dependent Manner ◽  
Major Pathway ◽  
Interstrand Crosslinks ◽  
Interstrand Crosslink ◽  
Pathway Activation ◽  
Base Excision ◽  
And Function

Abstract The NEIL3 DNA glycosylase is a base excision repair enzyme that excises bulky base lesions from DNA. Although NEIL3 has been shown to unhook interstrand crosslinks (ICL) in Xenopus extracts, how NEIL3 participants in ICL repair in human cells and its corporation with the canonical Fanconi anemia (FA)/BRCA pathway remain unclear. Here we show that the NEIL3 and the FA/BRCA pathways are non-epistatic in psoralen-ICL repair. The NEIL3 pathway is the major pathway for repairing psoralen-ICL, and the FA/BRCA pathway is only activated when NEIL3 is not present. Mechanistically, NEIL3 is recruited to psoralen-ICL in a rapid, PARP-dependent manner. Importantly, the NEIL3 pathway repairs psoralen-ICLs without generating double-strand breaks (DSBs), unlike the FA/BRCA pathway. In addition, we found that the RUVBL1/2 complex physically interact with NEIL3 and function within the NEIL3 pathway in psoralen-ICL repair. Moreover, TRAIP is important for the recruitment of NEIL3 but not FANCD2, and knockdown of TRAIP promotes FA/BRCA pathway activation. Interestingly, TRAIP is non-epistatic with both NEIL3 and FA pathways in psoralen-ICL repair, suggesting that TRAIP may function upstream of the two pathways. Taken together, the NEIL3 pathway is the major pathway to repair psoralen-ICL through a unique DSB-free mechanism in human cells.

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