key: cord-348104-7662q8dg
authors: Yang, Xiaoyun; Ma, Yinliang; Li, Yimiao; Dong, Yating; Yu, Lily L.; Wang, Hong; Guo, Lulin; Wu, Chen; Yu, Xiaochun; Liu, Xiuhua
title: Molecular basis for the MacroD1-mediated hydrolysis of ADP-ribosylation
date: 2020-06-22
journal: DNA Repair (Amst)
DOI: 10.1016/j.dnarep.2020.102899
sha: 
doc_id: 348104
cord_uid: 7662q8dg

MacroD1 is an enzyme that hydrolyzes protein mono-ADP-ribosylation. However, the key catalytic residues of MacroD1 in these biochemical reactions remain elusive. Here, we present the crystal structure of MacroD1 in a complex with ADP-ribose (ADPR). The β5-α10-loop functions as a switch loop to mediate substrate recognition and right orientation. The conserved Phe(272) in the β5-α10-loop plays a crucial role in the orientation of ADPR distal ribose, and a conserved hydrogen-bond network contributes significantly to hold and orient the catalytic water12, which mediates ADPR hydrolysis. Moreover, we found that MacroD1 was recruited to the sites of DNA damage via recognition of ADP-ribosylation at DNA lesions. The MacroD1-mediated ADPR hydrolysis is essential for DNA damage repair. Taken together, our study provides structural and functional insights into the molecular mechanism of MacroD1-mediated ADPR hydrolysis and its role in DNA damage repair.

that these DNA repair factors can be recycled to repair other lesions. If ADPR hydrolysis is suppressed, ADPR-binding DNA repair factors will be trapped at DNA lesions, which suppresses DNA damage repair. Thus, it has been shown that PARG is involved in both DNA single-strand break (SSB) repair and double-strand break (DSB) repair (8, 31) . TARG1 and ARH3 are also known to participate in SSB repair (8) . However, the role of MacroD1 in DNA damage repair has not been studied yet.

Although it has been shown that an N-terminal-truncated isoform of MacroD1 (residues N78-C325) localizes in mitochondria, alternatively the full-length isoform of MacroD1 (residues N1-C325) localizes in nucleus, especially under cellular stress, indicating that MacroD1 may play roles in multiple biological processes (9, 32) .

To understand the molecular mechanism of MacroD1-mediated ADPR hydrolysis, we determined the crystal structure of MacroD1 in a complex with ADPR. Our analyses reveal the detailed catalytic pocket of MacroD1 and shed light on its ADPR recognition and the molecular mechanism of ADPR hydrolysis. Moreover, our results demonstrate that MacroD1 is recruited to DNA lesions through the ADPribosylation recognition, and MacroD1-mediated ADPR hydrolysis plays a critical role in DNA damage repair. Based on the structural analyses, we characterized the key residues of MacroD1 that are involved in DNA damage repair.

To understand the catalytic mechanism of MacroD1, we determined the structure of the MacroD1-ADPR complex to 2.0 Å resolution using X-ray diffraction. The final model of the MacroD1-ADPR complex contains four protein molecules in the asymmetric unit, termed A, B, C and D respectively. The electron density map shows J o u r n a l P r e -p r o o f 6 that MacroD1 monomer binds to one ADPR molecule. The root mean square deviation (RMSD) between MacroD1 molecule A, B and C is less than 0.1 Å, revealing that the three MacroD1 molecules in the complex have nearly identical structure. The RMSD between molecule D and the other three is approximately 0.35 Å, which is mainly caused by the relative poor electron density of Molecule D. The MacroD1 molecule A combined with the ADPR ligand was used for the following structural analysis.

The MacroD1 monomer exhibits the canonical three-layered α-β-α sandwich with a central six-stranded β sheet containing a mixture of anti-parallel (β3-β4) and parallel (β2-β5-β6-β1) strands (Fig. 1A) . The ADPR molecule binds to the deep cleft of MacroD1 according to the 2Fo-Fc electron density map (Fig. 1B) . Structural alignment using the DALI server reveals the presence of many structural homologs of MacroD1 ( Fig. 1C and 1D (Fig. 1D ). All these proteins share common macro domain fold, which contains a three-layered α-β-α sandwich, and their ADPR molecules exhibit a similar conformation ( Fig. 1D and E) . Notably, the conformation of these macro domains are remarkably different from that of the ADPribosylhydrolase (ARH) family enzymes, including ARH1 (PDB code: 6IUX), DraG J o u r n a l P r e -p r o o f (PDB code: 2WOC) and ARH3 (PDB code: 5ZQY), which represent a compact all-αhelical fold.

Compared to apo-MacroD1 (PDB code: 2X47), the most distinctive structural characteristic in the MacroD1-ADPR complex is the dramatic conformational change of the β5-α10-loop ( Fig. 2A) . The loop covering the bound ADPR functions as a switch loop to sequester substrate and provides structural flexibility to accommodate alternative substrates ( Fig. 2A) indicating that the phenyl group is conserved in the macro domain hydrolase.

In addition, several conformational changes have been observed when ADPR binds to MacroD1. Val 271 from the β5-α10-loop was moved towards to the catalytic site about ~ 7Å (Fig. 2B) , and Phe 306 in β6-α11-loop undergoes a rotation to generate - stacking interaction with the adenine ring of ADPR (Fig. 2C ). Another important conformational change is to form the glycine-rich β2-α7-loop once recognizing ADPR. It is the part of α7 helix in the ligand-free state (PDB code: 2X47). The helix-J o u r n a l P r e -p r o o f 8 to-loop transition offers structural flexibility for the ADPR binding (Fig. 2D ).

ADPR is located at the deep substrate-binding groove of MacroD1. The ADPR molecule adopts an almost L-shaped conformation, and the long side consists of both pyrophosphate moiety and distal ribose of ADPR, and the adenosine moiety is loaded at the short side.

All the three parts of ADPR have extensive contacts with MacroD1 (Fig. 3A) . The adenine ring in the adenosine forms hydrogen-bonds with the side-chain carbonyl oxygen of Asp 160 and the main-chain amino group of Ile 161 . Furthermore, the adenine ring is also stacked by Phe 306 through the abovementioned - stacking interaction ( Fig. 2C ), which is a common characteristic for both macro domain hydrolases and ARH hydrolases (33) . The adenosine ribose forms hydrogen bonds with the sidechain of Thr 269 and three water molecules (water19, water90 and water155). The pyrophosphate moiety in ADPR is surrounded by the β2-α7-loop and β5-α10-loop, and the latter loop undergoes a significant conformational change upon ADPR binding ( Fig. 2A ). One residue (Val 183 ) from the β2-α7-loop, and four residues (Ser 268 , Gly 270 , Val 271 and Phe 272 ) from β5-α10-loop are directly involved in the interaction with diphosphate group through hydrogen-bonds (Fig. 3A) . The extensive network of hydrogen-bonds contributes to the majority of the binding energy of ADPR to MacroD1. The distal ribose locates to the catalytic site surrounded by the glycine-rich β2-α7-loop. Gly 182 in the β2-α7-loop contributes to anchor the distal ribose 1″-OH group, and a hydrogen-bond network is formed between Gly 182 , 1″-OH group of distal ribose, water12 and the α-phosphate of ADPR. Two hydrogen-bonds are formed J o u r n a l P r e -p r o o f between the 2″-OH group of distal ribose and Asn 174 (β2-α7-loop) and Asp 184 (helix α7) (Fig. 3A) . Sequence alignment results show that the ADPR interacting residues are highly conserved in the macro domain hydrolases (Fig. 1C) .

Additionally, structural comparison shows that the water12 is also conserved among the macro domain hydrolases, including the MacroD2-ADPR complex (PDB code: 4IQY), the YmdB-ADPR complex (PDB code: 5CB3) and the TbMDO-ADPR complex (PDB code: 5FSY) ( Fig. 3B-E) . The water12 lies close to the C1″ atom of the distal ribose with a hydrogen-bond distance of 3.1 Å. Based on the previous studies and our analysis on the MacroD1-ADPR complex, it suggests that the water12 is the catalytic water for the ADPR hydrolysis in MacroD1, and that the water12 will be activated and in turn launches a nucleophilic attack on the C1″ atom of the ADPR distal ribose.

In order to validate the structural analysis above, we generated 3 mutants in the ADPR binding pocket, including D160A, G182E and G182P. The G182P and G182E mutants introduce larger side chains, which will generate a steric hindrance and block the entrance of ADPR distal ribose, and the D160A mutant disrupts the hydrogenbond between ADPR and MacroD1. Consequently, The D160A, G182P and G182E mutations abolished the enzymatic activity of MacroD1-mediated ADPR hydrolysis ( Fig. S1 ). Consistently, the results from ITC assays show that G182P, G182E and D160A mutants cannot bind to ADPR (Fig. S2) . The corresponding residue for MacroD1 Val 271 is Ile 189 in MacroD2, the two residues are hydrophobic amino acids and the Val 271 main-chain, rather than its side-chain, forms a hydrogen-bond with the ADPR diphosphate group, so the V271I mutant retains the vast majority of enzymatic activity on ADPR hydrolysis (Fig. S1 ).

It has been suggested that Asn 174 , Asp 184 and His 188 are crucial residues for MacroD1 to deacetylate OAADPr, in which Asp 184 functions as the general base to activate the catalytic water molecule for deacetylation (21, 29) . The sequence alignment results show that these catalytically relevant residues are fully conserved among the MacroD1 homologs and its paralogs (Fig. 1C) . However, the D184A, N174A/D184A and D184A/H188A mutants almost retain the enzymatic activity of wild type MacroD1 on ADPR hydrolysis (Fig. 4A ), indicating that these residues are not essential for the MacroD1-mediated ADPR hydrolysis. Structural analysis reveals that the distance between Asp 184 and catalytic water12 is 4.9 Å, and there is no hydrogenbond formed between them, indicating that Asp 184 cannot act as the general base to activate the catalytic water12, which is in well agreement with our biochemical analyses. The function of Asp 184 and Asn 174 in the MacroD1-mediated ADPR hydrolysis may be only involved in the coordination of the distal ribose in ADPR.

Additionally, the distance between His 188 and the distal ribose in ADPR is almost 7 Å, indicating that it is not involved in the catalysis of MacroD1-mediated ADPR hydrolysis, which is also consistent with our ADPR hydrolyzation assays. Combining with our structural analysis and ADPR hydrolyzation assay, it suggests that distinct catalytic residues are responsible for the MacroD1-mediated ADPR hydrolysis, rather than the catalytic residues Asn 174 , Asp 184 and His 188 in the deacetylation of OAADPr.

It is observed that Phe 272 adopts a significant conformational change in the catalytic pocket of MacroD1 upon ADPR binding, and that the corresponding phenyl group is evolutionarily conserved among macro domain hydrolases. The phenylalanine corresponding residues in MacroD2, YmdB and TbMDO are Tyr 190 , Tyr 126 and Tyr 214 respectively (Fig. 1C) . These aromatic residues share very similar conformational J o u r n a l P r e -p r o o f orientation, and could provide a steric hindrance for the right orientation of ADPR distal ribose in the catalytic pocket ( Fig. 3B-3E ). As abovementioned, the ADPR molecules also adopt a similar orientation in the catalytic pocket (Fig. 1E ). In contrast, the corresponding residue of Phe 272 of MacroD1 is the Asn 316 in MacroH2A.1.1 macro domain, which is inactive towards ADPR hydrolysis. There is no effective steric hindrance occurring owing to the residue replacement, resulting in a distinct ADPR distal ribose conformation, in which the distal ribose exhibits a more extended conformation and its C1″ atom is far away from the structurally conserved water molecule (Fig. S3 ). Next, we generated the F272A mutant and measured its enzymatic activity on the ADPR hydrolysis in vitro. As predicted, the F272A mutation abolishes the enzymatic activity. However, this mutant protein still retains binding affinity with ADPR ( Fig. 4A and B) , indicating that Phe 272 plays an important role in the MacroD1-mediated ADPR hydrolysis, and the enzymatic activity impairment can be unrelated to the binding with ADPR. Hence, Phe 272 is crucial for the precise orientation of distal ribose in the catalytic pocket of MacroD1.

The crystal structure of MacroD1-ADPR complex has revealed that a hydrogenbond network is formed among Gly 182 , 1″-OH group of the distal ribose, catalytic water12, ADPR α-phosphate, water76, and Val 271 (Fig. 3A) . This hydrogen-bond network is also observed in other macro domain hydrolases and contributes significantly to the position of catalytic water12 at the catalytic site ( Fig. 3B-E) . The distance between water12 and α-phosphate of ADPR is 2.7 Å, and The distance between water12 and the C1″ atom of the ADPR distal ribose is 3.1 Å. Considering the activity of ADPR α-phosphate in a polar environment and the substrate-assisted catalysis in which the functional group from a substrate is involved in the catalytic activity of its corresponding enzyme (34) , it suggests that the ADPR α-phosphate J o u r n a l P r e -p r o o f functions as general base to activate water12, which induces a nucleophilic attack to the C1″ atom of the ADPR distal ribose (Fig. 4C) .

Collectively, we propose a biochemical reaction mechanism for the MacroD1mediated ADPR hydrolysis. Upon ADPR binding, the Phe 272 in the β5-α10-loop flips into the catalytic pocket and keeps the ADPR distal ribose in the right orientation through steric hindrance effect and dipole interaction. Meanwhile, a hydrogen-bond network, linking water12, ADPR α-phosphate and other elements, is responsible for the precise position of water12 in the catalytic pocket. The water12 is then activated by the ADPR α-phosphate and induces a nucleophilic attack to the distal ribose C1″ atom of ADPR, cleaving the glycosidic bond between the distal ribose C1″ atom and the acceptor Asp/Glu residue. The significance of ADPR α-phosphate and the catalytic water in the MacroD2-mediated ADPR hydrolysis have been reported by Gytis Jankevicius et al (22) . Collectively, the evolutionarily conserved aromatic residue, structurally conserved catalytic water and the same conformation of ADPR in the catalytic pocket suggest that the MacroD-like macro domain hydrolases may use the same catalytic mechanism for the ADPR hydrolysis.

The enzymatic activity of MacroD1 plays an important role in DNA damage repair ADPR hydrolases are known to regulate DNA damage repair. Although MacroD1 is reported to exist in mitochondria, it has been shown that similar to its paralog MacroD2, at least a small amount of MacroD1 localizes in nucleus, particularly relocalizes to nucleus in response to cellular stress (9, 22) . Using laser microirradiation assays, we examined the recruitment of MacroD1 to DNA lesions. MacroD1 was recruited to the DNA lesions within 30 seconds following laser-induced DNA J o u r n a l P r e -p r o o f damage, and this relocation process was suppressed by the PARP inhibitor olaparib treatment (Fig. 5A) , suggesting that the recruitment of MacroD1 to DNA lesions is mediated by DNA damage-induced ADP-ribosylation. Compared to wild type MacroD1, D160A, G182P and G182E mutants failed to relocate to DNA lesions since these mutants disrupted the ADPR-binding pocket, indicating that ADPR recognition plays a key role for the recruitment of MacroD1 to DNA damage site (Fig. 5B) .

Interestingly, the F272A mutant was still recruited to DNA damage site (Fig. 5C) , suggesting that the catalytic-related residue is not required for the recruitment of MacroD1 to DNA damage site. Thus, these results reveal that ADPR recognition is essential for the recruitment of MacroD1 to DNA damage sites.

Next, to examine the role of MacroD1 in DNA damage repair, we knocked down the endogenous MacroD1 with siRNA, and expressed wild type MacroD1 and its mutants in U2OS cells (Fig. 6A) . The transfected cells were treated with methyl methanesulfonate (MMS), a DNA damaging reagent. The DNA damage repair kinetics was measured using comet assays. Under both neutral and alkaline conditions, the DNA damage repair was impaired once cells were only expressing MacroD1 mutants ( Fig. 6B and C) , suggesting that MacroD1-mediated ADPR hydrolyzation plays a crucial role in DNA damage repair.

In this study, we have solved the structure of the MacroD1-ADPR complex, which allows us to dissect the molecular mechanism of MacroD1-mediated ADPR hydrolysis. Although the crystal structure of apo-MacroD1 has been solved (21), the true catalytic residues for the ADPR hydrolysis were unclear until this study. (29) . They proposed that Asp 184 works as a general base to deprotonate the catalytic water, which further launches an nucleophilic attack on the carbonyl group, resulting in the hydrolyzation of the acetyl group from OAADPr.

In this study, compared with apo-MacroD1, our structure of MacroD1-ADPR complex reveals the conformational plasticity of β5-α10-loop, which exists in open and closed conformational states. A structural rearrangement, required for the specific substrate recognition, is observed upon ADPR binding (Fig. 2) . Additionally, the structural conserved water (water12) is observed in the catalytic site of MacroD1, which also existed in other MacroD-like proteins (Fig. 3) TbMDO-ADPR complex (PDB code: 5FSY) (Fig. 3B-E) . Additionally, their ADPR molecules adopt a similar conformation. Interestingly, several strains of virus, such as coronaviruses, also contain MacroD-like domains that retain the conserved Phe residues at the similar positions (Fig. S4) , suggesting that these macro domains may also regulates ADPR hydrolysis during viral infection (35) (36) (37) . Both evolutionary conservation and conformational similarity of the phenyl group indicate that the significance of aromatic residues in the ADPR hydrolysis. In contrast, the structure of ADPR bound to the MacroH2A1.1 macro domain (inactive) reveals that the corresponding residue for MacroD1 Phe 272 is Asn 316 , and the disappearance of steric hindrance, which is generated by phenyl group, makes the distal ribose in a relatively extended conformation, in which its C1″ atom is far away from the catalytic water ( Fig. S3) (38) . It is assumed that the presence of phenyl group plays a significant role in catalysis process. Indeed, it has been reported that the YmdB Y126A mutation completely abolishes the enzymatic activity on ADPR hydrolysis, whereas Y126F mutant had little impact on its enzymatic activity (28) . Taken together, it is suggested that the evolutionally conserved phenyl group from aromatic amino acid plays an J o u r n a l P r e -p r o o f important role on the ADPR hydrolysis, Phe 272 can provide the right orientation and stabilization of ADPR distal ribose in the catalytic site. More importantly, it seems that it is an efficient evolution strategy to replace a single amino acid to make the catalytic components in right orientation to complete the desired function.

Structure comparison also reveals that a hydrogen-bond network, linking the catalytic water, ADPR α-phosphate and other residues, is conserved in the macro domain hydrolases (Fig. 3B-E) . The common hydrogen-bond network contributes significantly to hold and orient catalytic water in the catalytic site. Both structural analysis and biochemical analysis have shown that Asp 184 cannot work as the general base to activate the catalytic water at the process of ADPR hydrolysis. The absence of a residue to activate catalytic water in the catalytic site and the substrate-assisted catalysis mechanism allow us to surmise that the ADPR α-phosphate works as the general base to activate the catalytic water12, which further carries on a nucleophilic attack on the distal ribose C1″ atom. The phenomenon that the involvement of a substrate phosphate as a general base in the enzymatic activity has been observed in many other enzymes, such as restriction endonucleases, aminoacyl tRNA syanthetases, RNase HI, aspartate carbamoyl-transferase, and so on (34) .

In addition to the structural and biochemical analysis, we further explored the roles of abovementioned residues in the MacroD1-mediated DNA damage repair. Laser microirradiation assays revealed that MacroD1 is recruited to DNA lesions, which is mediated by DNA damage-induced PARylation (Fig. 5A) . The ADPR binding residues are required for the recruitment of MacroD1 to DNA lesions (Fig. 5B) .

However, Phe 272 , although very important for catalytic activity of ADPR hydrolysis, is not necessary for the recruitment of MacroD1 to DNA damage site, indicating that the catalytic-related residue is not responsible for the recruitment of MacroD1 to J o u r n a l P r e -p r o o f DNA lesions (Fig. 5C ). Furthermore, due to the lack of enzymatic activity, the MacroD1 mutants abolished the MacroD1-mediated DNA damage repair (Fig. 6) .

Accumulated evidence suggests that ADP-ribosylation mediates the recruitment of DNA damage repair factors to the DNA damage sites for lesion repair, and deADPribosylation is a sequential step to remove ADP-ribosylation and facilitates repair. If deADP-ribosylation is abolished, these DNA damage repair factors will be trapped at DNA lesions by ADP-ribosylation and impair DNA damage repair (7, 8) . It has been shown that dePARylation plays a key role in DNA damage repair (8, 30) .

Consistently, we found that lacking MacroD1 also impaired DNA damage repair. In addition to MacroD1, PARG is another potent enzyme to remove PARylation during DNA damage repair. However, it cannot remove the last ADPR unit linked to the targeted proteins. In contrast, MacroD1 is able to remove the terminal ADPR unit In summary, our study provides mechanistic insights into the MacroD1-mediated ADPR hydrolysis, which may explain its biological functions in cellular processes such as DNA damage repair.

The DNA sequence encoding Human MacroD1 was amplified from a 293T cDNA library. The amplified PCR product was digested with BamHI and XhoI, and then was 

To screen the MacroD1-ADPR complex, MacroD1 and ADPR (Sigma) were mixed in the molar ratio of 1:3. Preliminary crystallization was performed using the sitting- Crystal diffraction data were collected at Shanghai Synchrotron Radiation facility (SSRF), beamline BL19U .The crystal belongs to space group P1121 with the unit cell dimensions a = 79.9 Å, b = 106.2 Å, c = 79.9 Å, and β = 120.00°. The data set was processed with XDS (39) .

The crystal structure of MacroD1-ADPR complex was determined through molecular replacement using PHASER from the CCP4 software package (40) . The structure of apo-MacroD1 (PDB code: 2X47) was used as the search model. COOT (41) and J o u r n a l P r e -p r o o f PHENIX (42) were used repeatedly for manual model building and refinement. The data collection and refinement statistics are summarized in Table 1 . All the molecular graphics figures are prepared using PyMol (http://www.pymol.org).

U2OS cells were maintained in DMEM medium with 10 % fetal bovine serum and cultivated at 37 °C in 5 % CO2 (v/v). U2OS cells were transfected with plasmids encoding MacroD1 or its mutants.

Human MacroD1 and its mutants were cloned into pEGFP-C1, pET-15b, and pCDNA3 vectors. Human PARP10 full-length cDNA was cloned to the pGEX-4T-1 vector. MacroD1 siRNA targeting sequence was 5' -GGAGCCCAGGUAUAAAAAGUU-3'.

The transfected U2OS Cells with GFP-tagged corresponding plasmids were plated on 

We performed single-cell gel electrophoretic comet assay to detect single strand DNA breaks under alkaline condition and DNA double strand breaks under neutral conditions. U2OS cells were recovered in normal culture medium for a certain period of time with or without the indicated treatment. Cells were collected and washed twice with ice-cold PBS; 2 × 10 4 /mL cells were mixed with 1 % LMAgarose at a ratio 

The auto-ADP-ribosylation of PARP10 was used as the substrate of MacroD1 in the ADPR hydrolysis assay and prepared using the previously published methods (31) . 

Biology of Poly(ADP-Ribose) Polymerases: The Factotums of Cell Maintenance

Structures and Mechanisms of Enzymes Employed in the Synthesis and Degradation of PARP-Dependent Protein ADP-Ribosylation

The PARP superfamily

Functional aspects of protein mono-ADP-ribosylation

Poly(ADP-ribose): an organizer of cellular architecture

Family-wide analysis of poly(ADP-ribose) polymerase activity

The role of poly ADP-ribosylation in the first wave of DNA damage response

The role of dePARylation in DNA damage repair and cancer suppression

MacroD1 Is a Promiscuous ADP-Ribosyl Hydrolase Localized to 10

Identification and characterization of a mammalian 39-kDa poly(ADP-ribose) glycohydrolase

The 39-kDa poly(ADPribose) glycohydrolase ARH3 hydrolyzes O-acetyl-ADP-ribose, a product of the Sir2 family of acetyl-histone deacetylases

Serine ADP-ribosylation reversal by the hydrolase ARH3. Elife

Reversibility of argininespecific mono(ADP-ribosyl)ation: identification in erythrocytes of an ADPribose-L-arginine cleavage enzyme

The macro domain protein family: structure, functions, and their potential therapeutic implications

The macro domain is an ADPribose binding module

Macrodomains: Structure, Function, Evolution, and Catalytic Activities

Poly(ADP-ribose) catabolism in mammalian cells exposed to DNA-damaging agents

The recognition and removal of cellular poly(ADP-ribose) signals

Isolation and characterization of the cDNA encoding bovine poly(ADP-ribose) glycohydrolase

The structure and catalytic mechanism of a poly(ADP-ribose) glycohydrolase

Identification of macrodomain proteins as novel O-acetyl-ADP-ribose deacetylases

A family of macrodomain proteins reverses cellular mono-ADP-ribosylation

Orphan macrodomain protein (human C6orf130) is an O-acyl-ADP-ribose deacylase: solution structure and catalytic properties

Macrodomain-containing proteins are new mono-ADPribosylhydrolases

Deficiency of terminal ADP-ribose protein glycohydrolase TARG1/C6orf130 in neurodegenerative disease

Structural insights into the mechanism of Escherichia coli YmdB: A 2'-Oacetyl-ADP-ribose deacetylase

Transition-state analysis of 2-O-acetyl-ADP-ribose hydrolysis by human macrodomain 1

Targeting dePARylation for cancer therapy

Targeting dePARylation selectively suppresses DNA repair-defective and PARP inhibitor-resistant malignancies

Differential activities of cellular and viral macro domain proteins in binding of ADP-ribose metabolites

Structurefunction analyses reveal the mechanism of the ARH3-dependent hydrolysis of ADP-ribosylation

Substrate-assisted catalysis: molecular basis and biological significance

Structural and functional basis for ADP-ribose and poly(ADPribose) binding by viral macro domains

Crystal structures of two coronavirus ADP-ribose-1''-monophosphatases and their complexes with ADP-Ribose: a systematic structural analysis of the viral ADRP domain

Macro Domain from Middle East Respiratory Syndrome Coronavirus (MERS-CoV) Is an Efficient ADP-ribose Binding Module: CRYSTAL STRUCTURE AND BIOCHEMICAL STUDIES

A macrodomain-containing histone rearranges chromatin upon sensing PARP1 activation

Phaser crystallographic software

Features and development of Coot

PHENIX: a comprehensive Python-based system for macromolecular structure solution

The authors thank the beamline BL19U staff at the Shanghai Synchrotron Radiation