2017-2020 Publications
2017-2020 Publications
2017-2020 Publications
2017-Present Publications
Highlights
* = co-first author, † = corresponding author
Small-molecule GSDMD agonism in tumors stimulates antitumor immunity without toxicity
Fontana P*, Du G*, Zhang Y*, Zhang H*, Vora SM, Hu JJ, Shi M, Tufan AB, Healy LB, Xia S, Lee DJ, Li Z, Baldominos P, Ru H, Luo HR, Agudo J, Lieberman J.†, Wu H..†.. Cell (2024). PDF
Gasdermin-mediated inflammatory cell death (pyroptosis) can activate protective immunity in immunologically cold tumors. Here, we performed a high-throughput screen for compounds that could activate gasdermin D (GSDMD), which is expressed widely in tumors. ... Read More
NINJ1 mediates plasma membrane rupture by cutting and releasing membrane disks
David L*, Borges JP, Hollingsworth LR, Volchuk A, Jansen I, Garlick E, Steinberg BE, Wu H.†. Cell (2024) 187(9):2224-2235 PDF
The membrane protein NINJ1 mediates plasma membrane rupture in pyroptosis and other lytic cell death pathways. Here, we report the cryo-EM structure of a NINJ1 oligomer segmented from NINJ1 rings. Each NINJ1 subunit comprises amphipathic (⍺1, ⍺2) and transmembrane (TM) helices (⍺3, ⍺4) and forms a chain of subunits.... Read More
Dong Y*, Bonin JP, Devant P, Liang Z, Sever AIM, Mintseris J, Aramini JM, Du G, Gygi SP, Kagan JC, Kay LE, Wu H. †. Immunity ( 2024 ) May 8:S1074-7613(24)00220-6. PDF
Several interleukin-1 (IL-1) family members, including IL-1β and IL-18, require processing by inflammasome-associated caspases to unleash their activities. Here, we unveil, by cryoelectron microscopy (cryo-EM), two major conformations of the complex between caspase-1 and pro-IL-18. One conformation is similar to the complex of caspase-4 and pro-IL-18, with interactions at both the active site and an exosite (closed conformation), and the other only contains interactions at the active site (open conformation)....Read More
ROS-dependent S-palmitoylation activates cleaved and intact gasdermin D
Du G*, Healy LB*, David L, Walker C, El-Baba TJ, Lutomski CA, Goh B, Gu B, Pi X, Devant P, Fontana P, Dong Y, Ma X, Miao R, Balasubramanian A, Puthenveetil R, Banerjee A, Luo HR, Kagan JC, Oh SF, Robinson CV, Lieberman J, Wu H†. Nature ( 2024 ) 630 ( 8016 ) 437-446. PDF
Gasdermin D (GSDMD) is the common effector for cytokine secretion and pyroptosis downstream of inflammasome activation and was previously shown to form large transmembrane pores after cleavage by inflammatory caspases to generate the GSDMD N-terminal domain (GSDMD-NT)1-10. Here we report that GSDMD Cys191 is S-palmitoylated and that palmitoylation is required for pore formation. S-palmitoylation, which does not affect GSDMD cleavage, is augmented by mitochondria-generated reactive oxygen species (ROS). ... Read More
Fu J*, Schroder K, Wu H †. Nat Rev Immunol ( 2024 ) Feb 19 doi: 10.1038/s41577-024-00995w. PDF
Inflammasomes are supramolecular complexes that form in the cytosol in response to pathogen-associated and damage-associated stimuli, as well as other danger signals that perturb cellular homoeostasis, resulting in host defence responses in the form of cytokine release and programmed cell death (pyroptosis). Inflammasome activity is closely associated with numerous human disorders,including rare genetic syndromes of autoinflammation, cardiovascular diseases, neurodegeneration and cancer.....Read More
Devant P*, Dong Y*, Mintseris J, Ma W, Gygi SP, Wu H †, Kagan JC †. Nature ( 2023 ) Dec;624(7991):451-459. PDF
Inflammatory caspases are key enzymes in mammalian innate immunity that control the processing and release of interleukin-1 (IL-1)-family cytokines1,2. Despite the biological importance, the structural basis for inflammatory caspase-mediated cytokine processing has remained unclear. To date, catalytic cleavage of IL-1-family members, including pro-IL-1β and pro-IL-18, has been attributed primarily to caspase-1 activities within canonical inflammasomes3.....Read More
Du G *, Zhao L, Zheng Y, Belfetmi A, Cai T, Xu B, Heyninck K, Van Den Heede K, Buyse MA, Fontana P, Bowman M, Lin LL, Wu H†, Chou JJ† Cell Res. (2023) 33 (2):131-146. PDF
Members of the tumor necrosis factor receptor superfamily (TNFRSF) are important therapeutic targets that can be activated to induce death of cancer cells or stimulate proliferation of immune cells. Although it has long been implicated that these receptors assemble preligand associated states that are required .....Read More
Li J*, Wang L*, Hahn Q, Nowak R , Viennet T, Orellana E, Burman S, Yue H , Hunkeler M, Fontana P, Wu H, Arthanari H, Fischer E, Gregory R. Nature. (2023 ) Jan 4th, doi 10 1038/s41586- 022005566-4. PDF
Chemical modifications of RNA have key roles in many biological processes1-3. N7-methylguanosine (m7G) is required for integrity and stability of a large subset of tRNAs4-7. The methyltransferase 1-WD repeat-containing protein 4 (METTL1-WDR4) complex is the methyltransferase that modifies G46 in the variable loop of certain tRNAs, Read More
Cryo-EM structures of the active NLRP3 inflammasome disk
Inflammasomes are cytosolic innate immune complexes that activate caspase-1 upon detection of pathogenic and endogenous dangers1-5, and NLRP3 is an inflammasome sensor of membrane damage highly important in inducing inflammation2,6,7. Here we report cryo-EM structures of disk-shaped active NLRP3 oligomers in complex with ATP𝛾S, the centrosomal kinase NEK7, and the adaptor protein ASC which recruits caspase-1. In these NLRP3-NEK7-ASC complexes, the central NACHT domain of NLRP3 assumes an ATP-bound conformation in which two of its subdomains rotate by ~85 ° relative to the ADP-bound inactive conformation8-12. ... Read More
Structural principles of B cell antigen receptor assembly
Dong Y*, Pi X*, Bartels-Burgahn F, Saltukoglu D, Liang Z, Yang J, Alt FW, Reth M, Wu H † Nature (2022) , 612(7938):156-161. PDF
The B cell antigen receptor (BCR) is composed of a membrane-bound class M, D, G, E or A immunoglobulin for antigen recognition1–3 and a disulfde-linked Igα (also known as CD79A) and Igβ (also known as CD79B) heterodimer (Igα/β) that functions as the signalling entity through intracellular immunoreceptor tyrosine-based activation motifs (ITAMs)4,5 . The organizing principle of the BCR remains unknown.... Read More
Structure of cytoplasmic ring of nuclear pore complex by integrative cryo-EM and AlphaFold
Fontana P*, Dong Y*, Pi X*, Tong AB, Hecksel CW, Wang L, Fu TM, Bustamante C, Wu H †. Science. (2022) Jun 10;376(6598):eabm9326. doi: 10.1126/science.abm9326. PDF
The nuclear pore complex (NPC) is the molecular conduit in the nuclear membrane of eukaryotic cells that regulates import and export of biomolecules between the nucleus and the cytosol, with vertebrate NPCs ~110 to 125 MDa in molecular mass and ~120 nm in diameter. NPCs are organized into four main rings: the cytoplasmic ring (CR) at the cytosolic side, Read More
BCL10 Mutations Define Distinct Dependencies Guiding Precision Therapy for DLBCL
Xia M*, David L, Teater M, Gutierrez J, Wang X, Meydan C, Lytle A, Slack GW, Scott DW, Morin RD, Onder O, Elenitoba-Johnson KSJ, Zamponi N, Cerchietti L, Lu T, Philippar U, Fontan L, Wu H†, Melnick AM†. Cancer Discov. (2022) Aug 5;12(8):1922-1941. doi: 10.1158/2159-8290.CD-21-1566. PDF
Activated B cell-like diffuse large B-cell lymphomas (ABC-DLBCL) have unfavorable outcomes and chronic activation of CARD11-BCL10-MALT1 (CBM) signal amplification complexes that form due to polymerization of BCL10 subunits, which is affected by recurrent somatic mutations in ABC-DLBCLs. Read More
pH regulates potassium conductance and drives a constitutive proton current in human TMEM175
Zheng W*, Shen C, Wang L, Rawson S, Xie WJ, Nist-Lund C, Wu J, Shen Z, Xia S, Holt JR, Wu † Fu TM.. Sci Adv. (2022) Mar 25;8(12):eabm1568. doi: 10.1126/sciadv.abm1568. PDF
Human TMEM175, a noncanonical potassium (K+) channel in endolysosomes, contributes to their pH stability and is implicated in the pathogenesis of Parkinson's disease (PD). Structurally, the TMEM175 family exhibits an architecture distinct from canonical potassium channels, as it lacks the typical TVGYG selectivity filter. Read More
Targeting stem-loop 1 of the SARS-CoV-2 5' UTR to suppress viral translation and Nsp1 evasion
Vora SM*, Fontana P, Mao T, Leger V, Zhang Y, Fu TM, Lieberman J, Gehrke L, Shi M, Wang L, Iwasaki A, Wu H..†. Proc Natl Acad Sci U S A. (2022) Mar 1;119(9). doi: 10.1073/pnas.2117198119. PDF
SARS-CoV-2 is a highly pathogenic virus that evades antiviral immunity by interfering with host protein synthesis, mRNA stability, and protein trafficking. The SARS-CoV-2 nonstructural protein 1 (Nsp1) uses its C-terminal domain to block the messenger RNA (mRNA) entry channel of the 40S ribosome to inhibit host protein synthesis. However, how SARS-CoV-2 circumvents Nsp1-mediated suppression for viral protein synthesis Read More
Electrostatic influence on IL-1 transport through the GSDMD pore
Xie WJ*, Xia S, Warshel A, Wu H.†. Proc Natl Acad Sci U S A. (2022 ) Feb 8;119(6). doi10.1073/pnas.2120287119. PDF
A variety of signals, including inflammasome activation, trigger the formation of large transmembrane pores by gasdermin D (GSDMD). There are primarily two functions of the GSDMD pore, to drive lytic cell death, known as pyroptosis, and to permit the release of leaderless interleukin-1 (IL-1) family cytokines, a process that does not require pyroptosis. We are interested in the mechanism by which the GSDMD pore channels IL-1 release from living cells. Recent studies revealed that electrostatic interaction, in addition to cargo size, plays a critical role in GSDMD-dependent protein release. Read More
NLRP3 cages revealed by full-length mouse NLRP3 structure control pathway activation
Andreeva L*, David L, Rawson S, Shen C, Pasricha T, Pelegrin P, Wu H.†. Cell 184, 1–16 (2021). PDF
"The NACHT-, leucine-rich-repeat- (LRR), and pyrin domain-containing protein 3 (NLRP3) is emerging to be a critical intracellular inflammasome sensor of membrane integrity and a highly important clinical target against chronic inflammation. Here, we report that an endogenous, stimulus-responsive form of full-length mouse NLRP3 is a 12- to 16-mer double-ring cage held together by LRR-LRR interactions with the pyrin domains shielded within the assembly to avoid premature activation. Read More
Phase separation drives RNA virus-induced activation of the NLRP6 inflammasome
NLRP6 is important in host defense by inducing functional outcomes including inflammasome activation and interferon production. Here, we show that NLRP6 undergoes liquid-liquid phase separation (LLPS) upon interaction with double-stranded RNA (dsRNA) in vitro and in cells, and an intrinsically disordered poly-lysine sequence (K350-354) of NLRP6 is important for multivalent interactions, phase separation, Read More
Dipeptidyl peptidase 9 sets a threshold for CARD8 inflammasome formation by sequestering its active C-terminal fragment
Sharif H*, Hollingsworth LR*, Griswold AR*, Hsiao JC, Wang Q, Bachovchin DA†., Wu H† Immunity 54: 1-13 (2021). PDF
Inflammasomes are multiprotein complexes that execute pyroptosis in response to cytosolic danger signals. DPP9 regulates the CARD8 inflammasome by unknown mechanisms. Here, Sharif et al. report cryo-EM structures of CARD8 bound to DPP9 and show that this complex inhibits CARD8 inflammasome activation downstream of the proteasome. Read More
Gasdermin D pore structure reveals preferential release of mature interleukin-1
Xia S, Zhang Z, Magupalli VG, Pablo JL, Dong Y, Vora SM, Wang L, Fu T-M, Jacobson MP, Greka A, Lieberman J, Ruan J†,, Wu H†. Nature 593: 607-611 (2021) PDF
As organelles of the innate immune system, inflammasomes activate caspase-1 and other inflammatory caspases that cleave gasdermin D (GSDMD). Caspase-1 also cleaves inactive precursors of the interleukin (IL)-1 family to generate mature cytokines such as IL-1β and IL-18. Cleaved GSDMD forms transmembrane pores to enable the release of IL-1 and to drive cell lysis through pyroptosis. Here we report cryo-electron microscopy structures of the pore and the prepore of GSDMD. Read More
DPP9 sequesters the C terminus of NLRP1 to repress inflammasome activation
Hollingsworth, L.R.*, Sharif, H.*, Griswold, A.R.*, Fontana P., Mintseris J., Dagbay K.B., Paulo J.A., Gygi S.P., Bachovchin D.A.†, Wu H.† Nature 592, 778–783 (2021). PDF
Nucleotide-binding domain and leucine-rich repeat pyrin-domain containing protein 1 (NLRP1) is an inflammasome sensor that mediates the activation of caspase-1 to induce cytokine maturation and pyroptosis. Gain-of-function mutations of NLRP1 cause severe inflammatory diseases of the skin. NLRP1 contains a function-to-find domain that auto-proteolyses into noncovalently associated subdomains, and proteasomal degradation of the repressive N-terminal fragment of NLRP1 releases its inflammatory C-terminal fragment (NLRP1 CT). Read More
Mechanism of filament formation in UPA-promoted CARD8 and NLRP1 inflammasomes
Hollingsworth LR*., David L*., Li Y*., Griswold AR., Ruan J., Sharif H., Fontana P., Orth-He EL., Fu, T., Bachovchin DA, Wu H.† Nat Commun 12, 189 (2021). PDF
NLRP1 and CARD8 are related cytosolic sensors that upon activation form supramolecular signalling complexes known as canonical inflammasomes, resulting in caspase-1 activation, cytokine maturation and/or pyroptotic cell death. NLRP1 and CARD8 use their C-terminal (CT) fragments containing a caspase recruitment domain (CARD) and the UPA (conserved in UNC5, PIDD, and ankyrins) Read More
HDAC6 mediates an aggresome-like mechanism for NLRP3 and pyrin inflammasome activation
Venkat Giri Magupalli*†, Roberto Negro*†, Yuzi Tian*, Arthur V. Hauenstein*, Giuseppe Di Caprio, Wesley Skillern, Qiufang Deng, Pontus Orning, Hasan B. Alam, Zoltan Maliga, Humayun Sharif, Jun Jacob Hu, Charles L. Evavold, Jonathan C. Kagan, Florian I. Schmidt, Katherine A. Fitzgerald, Tom Kirchhausen, Yongqing Li†, Hao Wu†. Science (2020). PDF
Inflammasomes are supramolecular complexes that play key roles in immune surveillance. This is accomplished by the activation of inflammatory caspases, which leads to the proteolytic maturation of interleukin 1β (IL-1β) and pyroptosis. Here, we show that nucleotide-binding domain, leucine-rich repeat, . Read More
FDA-approved disulfiram inhibits pyroptosis by blocking gasdermin D pore formation
Hu JJ*, Liu X*†, Xia S, Zhang Z, Zhang Y, Zhao J, Ruan J, Luo X, Lou X, Bai Y, Wang J, Hollingsworth LR, Magupalli VG, Zhao L, Luo HR, Kim J, Lieberman J†, Wu H†.. Nature Immunol (2020). PDF
Cytosolic sensing of pathogens and damage by myeloid and barrier epithelial cells assembles large complexes called inflammasomes, which activate inflammatory caspases to process cytokines (IL-1β) and gasdermin D (GSDMD). Cleaved GSDMD forms membrane pores, Read More
Gasdermin D activity in inflammation and host defense
Lieberman J*†, Wu H*†, Kagan JC*†. Sci Immunol (2019). PDF
The mechanisms underlying the release of interleukin-1 (IL-1) family cytokines from phagocytes have been the subject of intense investigations for more than 30 years. The absence of an amino-terminal secretion signal from members of this family suggests a previously unknown mechanism of protein secretion that transfers cytosolic IL-1 directly across the plasma membrane into the extracellular space. The pore-forming protein gasdermin D (GSDMD) has emerged as the conduit for... Read More
Structural Mechanism for NEK7-licensed activation of NLRP3 inflammasome
Sharif H*, Wang L*, Wang WL*, Magupalli VG, Andreeva L, Qiao Q, Hauenstein AV, Wu Z, Núñez G, Mao Y†, Wu H†. Nature (2019). PDF
The NLRP3 inflammasome can be activated by stimuli that include nigericin, uric acid crystals, amyloid-β fibrils and extracellular ATP. The mitotic kinase NEK7 licenses the assembly and activation of the NLRP3 inflammasome in interphase. Here we report a cryo-electron microscopy structure of inactive human NLRP3 in complex with NEK7, at a resolution of 3.8 Å. Read More
Higher-Order Clustering of the Transmembrane Anchor of DR5 Drives Signalling
Pan L*, Fu TM*, Zhao W*, Zhao L*, Chen W, Qiu C, Liu W, Liu Z, Piai A, Fu Q, Chen S†, Wu H†, Chou JJ†. Cell (2019). PDF
Receptor clustering on the cell membrane is critical in the signaling of many immunoreceptors, and this mechanism has previously been attributed to the extracellular and/or the intracellular interactions. Here, we report an unexpected finding that for death receptor 5 (DR5), a receptor in the tumor necrosis factor receptor superfamily, the transmembrane helix (TMH) alone in the receptor directly.. Read More
Crystal structure of the WD40 domain dimer of LRRK2
Zhang P*, Fan Y*, Ru H, Wang L, Magupalli VG, Taylor SS, Alessi DR, Wu H*†. Proc Natl Acad Sci U S A (2019). PDF
Leucine-rich repeat kinase 2 (LRRK2) is a large multidomain protein with both a Ras of complex (ROC) domain and a kinase domain (KD) and, therefore, exhibits both GTPase and kinase activities. Human genetics studies have linked LRRK2 as a major genetic contributor to familial and sporadic Parkinson’s disease (PD), a neurodegenerative movement disorder that inflicts millions worldwide. Read More
Molecular mechanism for NLRP6 inflammasome assembly and activation
Shen C*, Lu A, Xie WJ, Ruan J, Negro R, Egelman EH, Fu TM*†, Wu H*†. Proc Natl Acad Sci U S A (2019). PDF
Inflammasomes are large protein complexes that trigger host defense in cells by activating inflammatory caspases for cytokine maturation and pyroptosis. NLRP6 is a sensor protein in the nucleotide-binding domain (NBD) and leucine-rich repeat (LRR)-containing (NLR) inflammasome family that has been shown to play multiple roles in regulating inflammation and host defenses. Despite the significance of the NLRP6 inflammasome, little is known about the molecular mechanism behind... Read More
Conformational flexibility and inhibitor binding to unphosphorylated interleukin-1-receptor-associated kinase 4 (IRAK4)
Wang L*, Ferrao R*, Li Q, Hatcher JM, Choi HG, Buhrlage SJ, Gray NS, Wu H†. The Journal of Biological Chemistry (2019). PDF
Interleukin-1 receptor–associated kinase 4 (IRAK4) is a key player in innate immune and inflammatory responses, performing a critical role in signal transduction downstream of Toll-like receptors and interleukin-1 (IL-1) receptors. Upon ligand binding and via its N-terminal death domain, IRAK4 is ... Read More
2021
Cryo-EM structures of ASC and NLRC4 CARD filaments reveal a unified mechanism of nucleation and activation of caspase-1
Li Y*, Fu TM*†, Lu A, Witt K, Ruan J, Shen C, Wu H†. Proc Natl Acad Sci U S A (2018). PDF
Canonical inflammasomes are cytosolic supramolecular complexes that activate caspase-1 upon sensing extrinsic microbial invasions and intrinsic sterile stress signals. During inflammasome assembly, adaptor proteins ASC and NLRC4 recruit caspase-1 through homotypic caspase recruitment domain (CARD) interactions, leading to caspase-1 dimerization and activation. Activated caspase-1 processes...Read More
Structures and gating mechanism of human TRPM2
Transient receptor potential (TRP) melastatin 2 (TRPM2) is a cation channel associated with numerous diseases. It has a C-terminal NUDT9 homology (NUDT9H) domain responsible for binding adenosine diphosphate (ADP)–ribose (ADPR), and both ADPR and calcium (Ca2+) are required for TRPM2 activation. Here we report cryo–electron microscopy structures of human TRPM2 alone, with ADPR, and with ADPR and Ca2+. NUDT9H forms both intra- and intersubunit interactions with the N-terminal TRPM homology... Read More
DNA melting initiates the RAG catalytic pathway
Ru H*, Mi W*, Zhang P, Alt FW, Schatz DG, Liao M†, Wu H †. Nature structural & molecular biology (2018). PDF
The mechanism for initiating DNA cleavage by DDE-family enzymes, including the RAG endonuclease, which initiates V(D)J recombination, is not well understood. Here we report six cryo-EM structures of zebrafish RAG in complex with one or two intact recombination signal sequences (RSSs), at up to 3.9-Å resolution. Unexpectedly, these structures reveal DNA melting at the heptamer of the RSSs, thus resulting in a corkscrew-like rotation of coding-flank DNA and the positioning of the scissile phosphate in the active site. Read More
The Structure of the Necrosome RIPK1-RIPK3 Core, a Human Hetero-Amyloid Signaling Complex
The RIPK1-RIPK3 necrosome is an amyloid signaling complex that initiates TNF-induced necroptosis, serving in human immune defense, cancer, and neurodegenerative diseases. RIPK1 and RIPK3 associate through their RIP homotypic interaction motifs with consensus sequences IQIG (RIPK1) andVQVG (RIPK3). Using solid-state nuclear magnetic resonance, we determined the high-resolution structure of the RIPK1-RIPK3 core. Read More
Cryo-EM structure of the gasdermin A3 membrane pore
Ruan J, Xia S, Liu X, Lieberman J, Wu H†. (2018). PDF
Gasdermins mediate inflammatory cell death after cleavage by caspases or other, unknown enzymes. The cleaved N-terminal fragments bind to acidic membrane lipids to form pores, but the mechanism of pore formation remains unresolved. Here we present the cryo-electron microscopy structures of the 27-fold and 28-fold single-ring pores formed by the N-terminal fragment of mouse GSDMA3 (GSDMA3-NT) at 3.8 and 4.2Å resolutions, and of a double-ring pore at 4.6Å resolution. In the 27-fold pore, a 108-stranded... Read More
Assembly mechanism of the CARMA1-BCL10-MALT1-TRAF6 signalosome
David L, Li Y, Ma J, Garner E, Zhang X, Wu H†. Proc Natl Acad Sci U S A (2018). PDF
The CARMA1–BCL10–MALT1 (CBM) signalosome is a central mediator of T cell receptor and B cell receptor-induced NF-κB signaling that regulates multiple lymphocyte functions. While caspaserecruitment domain (CARD) membrane-associated guanylate kinase (MAGUK) protein 1 (CARMA1) nucleates B cell lymphoma 10 (BCL10) filament formation through interactions between CARDs, mucosaassociated lymphoid tissue lymphoma translocation protein 1 (MALT1) is a paracaspase with structural similarity to caspases, Read More
Crystal structure of human IRAK1
Wang L*, Qiao Q, Ferrao R*, Shen C, Hatcher JM, Buhrlage SJ, Gray NS, Wu H†. Proc Natl Acad Sci U S A (2017). PDF
Interleukin 1 (IL-1) receptor-associated kinases (IRAKs) are serine/threonine kinases that play critical roles in initiating innate immune responses against foreign pathogens and other types of dangers through their role in Toll-like receptor (TLR) and interleukin 1 receptor (IL-1R) mediated signaling pathways. Upon ligand binding, TLRs and IL-1Rs recruit adaptor proteins, such as myeloid differentiation primary response gene 88 (MyD88), to the membrane, which in turn recruit IRAKs via the death domains... Read More
AID Recognizes Structured DNA for Class Switch Recombination
Activation-induced cytidine deaminase (AID) initiates both class switch recombination (CSR) and somatic
hypermutation (SHM) in antibody diversification. Mechanisms of AID targeting and catalysis remain elusive despite its critical immunological roles and off-target effects in tumorigenesis. Here, we produced active human AID and revealed its preferred recognition and deamination of structured substrates. G-quadruplex (G4)-containing substrates mimicking the mammalian immunoglobulin switch regions are... Read More
CIDE domains form functionally important higher-order assemblies for DNA fragmentation
Choi JY*, Qiao Q*, Hong SH, Kim CM, Jeong JH, Kim YG, Jung YK, Wu H†, Park HH†. Proc Natl Acad Sci U S A (2017). PDF
Cell death-inducing DFF45-like effector (CIDE) domains, initially identified in apoptotic nucleases, form a family with diverse functions ranging from cell death to lipid homeostasis. Here we show that the CIDE domains of Drosophila and human apoptotic nucleases Drep2, Drep4, and DFF40 all form head-to-tail helical filaments. Read More
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Cryo-EM structure of the DNA-PK holoenzyme
Sharif H*, Li Y*, Dong Y, Dong L, Wang WL, Mao Y, Wu H†. Proc Natl Acad Sci U S A (2017). PDF
DNA-dependent protein kinase (DNA-PK) is a large protein complex central to the nonhomologous end joining (NHEJ) DNA-repair pathway. It comprises the DNA-PK catalytic subunit (DNA-PKcs) and the heterodimer of DNA-binding proteins Ku70 and Ku80. Here, we report the cryo-electron microscopy (cryo-EM) structures of human DNA-PKcs at 4.4-Å resolution and the DNA-PK holoenzyme at 5.8-Å resolution. The DNA-PKcs structure contains three distinct segments: the N-terminal region with an arm and a bridge... Read More
Other Publications (2017-Present)
* = co-first author, † = corresponding author
Yan Y, Xiao J, Huang F, Xian W, Yu B, Cheng R, Wu H, Lu X, Wang X, Huang W, Li J, Oyejobi GK, Robinson CV, Wu H, Wu D, Liu X, Wang L, Zhu B .Phage defence system CBASS is regulated by a prokaryotic E2 enzyme that imitates the ubiquitin pathway. Nat Microbiol. 2024;9(6):1566-1578..
Tan T, Gao B, Yu H, Pan H, Sun Z, Lei A, Zhang L, Lu H, Wu H, Daley GQ, Feng Y, Zhang J Dynamic nucleolar phase separation influenced by non-canonical function of LIN28A instructs pluripotent stem cell fate decisions. . Nat Commun. 2024;15(1):1256.
Li R, Zan Y, Wang D, Chen X, Wang A, Tan H, Zhang G, Ding S, Shen C, Wu H, Zhu S..† A mouse model to distinguish NLRP6-mediated inflammasome-dependent and -independent functions.. Proc Natl Acad Sci U S A. 2024;121(6):e2321419121.
Wang Y, Hollingsworth LR, Sangaré LO, Paredes-Santos TC, Krishnamurthy S, Penn BH, Wu H, Saeij JPJ. I, Host E3 ubiquitin ligase ITCH mediates Toxoplasma gondii effector GRA35-triggered NLRP1 inflammasome activation and cell-autonomous immunity.
mBio. 2024 15 (3) e0330223.
Balasubramanian A, Hsu AY, Ghimire L, Tahir M, Devant P, Fontana P, Du G, Liu X, Fabin D, Kambara H, Xie X, Liu F, Hasegawa T, Xu R, Yu H, Chen M, Kolakowski S, Trauger S, Larsen MR, Wei W, Wu H, Kagan JC, Lieberman J, Luo HR The palmitoylation of gasdermin D directs its membrane translocation and pore formation during pyroptosis. Sci Immunol. 2024;9(94):eadn1452.
Miao R, Jiang C, Chang WY, Zhang H, An J, Ho F, Chen P, Zhang H, Junqueira C, Amgalan D, Liang FG, Zhang J, Evavold CL, Hafner-Bratkovič I, Zhang Z, Fontana P, Xia S, Waldeck-Weiermair M, Pan Y, Michel T, Bar-Peled L, Wu H, Kagan JC, Kitsis RN, Zhang P, Liu X, Lieberman J. Gasdermin D permeabilization of mitochondrial inner and outer membranes accelerates and enhances pyroptosis
Immunity 2023 Nov 1456 (11) 2523-2541 .
Healy LB*,Du G, Wu H.† Chopped! Newfound GSDMD cleavage facilitates tolerance to food allergens. Trends Immunol. 2023 Aug;44(8):571-573. doi: 10.1016/j.it.2023.06.006.
Kong Q, Xia S, Pan X, Ye K, Li Z, Li H, Tang X, Sahni N, Yi SS, Liu X, Wu H, Elowitz MB, Lieberman J, Zhang Z. Alternative splicing of GSDMB modulates killer lymphocyte-triggered pyroptosis. Sci Immunol. 2023 Apr 28;8(82):eadg3196. doi: 10.1126/sciimmunol.adg3196.
Sen Santara S, Lee DJ, Crespo Â, Hu JJ, Walker C, Ma X, Zhang Y, Chowdhury S, Meza-Sosa KF, Lewandrowski M, Zhang H, Rowe M, McClelland A, Wu H, Junqueira C, Lieberman J.† The NK cell receptor NKp46 recognizes ecto-calreticulin on ER-stressed cells Nature. 2023 Apr 5. doi: 10.1038/s41586-023-05912-0.
Baris S, Benamar M, Chen Q, Catak MC, Martínez-Blanco M, Wang M, Fong J, Massaad MJ, Sefer AP, Kara A, Babayeva R, Eltan SB, Yucelten AD, Bozkurtlar E, Cinel L, Karakoc-Aydiner E, Zheng Y, Wu H, Ozen A, Schmitz-Abe K, Chatila TA. Severe allergic dysregulation due to a gain of function mutation in the transcription factor STAT6. J Allergy Clin Immunol. 2023 Feb 8:S0091-6749(23)00151-3. doi: 10.1016/j.jaci.2023.01.023.
Fu J, Wu H.† Structural Mechanisms of NLRP3 Inflammasome Assembly and Activation Annu Rev Immunol. 2023 Vol. 41:301-316.
David L*, Martinez L, Xi Q, Kooshesh KA, Zhang Y, Shah JV, Maas RL†, Wu H.† Piezo mechanosensory channels regulate centrosome integrity and mitotic entry. Proc Natl Acad Sci U S A. 2023 Jan 3;120(1):e2213846120. doi: 10.1073/pnas.2213846120
Paidimuddala B, Cao J, Nash G, Xie Q, Wu H, Zhang L.. Mechanism of NAIP-NLRC4 inflammasome activation revealed by cryo-EM structure of unliganded NAIP5 . Nat Struct Mol Biol. 2023 Feb;30(2):159-166. doi: 10.1038/s41594-022-00889-2
Du T, Song Y, Ray A, Wan X, Yao Y, Samur MK, Shen C, Penailillo J, Sewastianik T, Tai YT, Fulciniti M, Munshi NC, Wu H, Carrasco RD, Ubiquitin receptor PSMD4/Rpn10 is a novel therapeutic target in multiple myeloma Chauhan D, Anderson KC. Blood. 2023 Jan 11:blood.2022017897. doi: 10.1182/blood.2022017897.
Baris S, Benamar M, Chen Q, Catak MC, Martínez-Blanco M, Wang M, Fong J, Massaad MJ, Sefer AP, Kara A, Babayeva R, Eltan SB, Yucelten AD, Bozkurtlar E, Cinel L, Karakoc-Aydiner E, Zheng Y, Wu H, Ozen A, Schmitz-Abe K, Chatila TA. Severe Allergic Dysregulation Due to A Gain of Function Mutation in the Transcription Factor STAT6.. J Intern Med. 2023 Mar;293(3):340-353.
Sun Z, Yu H, Zhao J, Tan T, Pan H, Zhu Y, Chen L, Zhang C, Zhang L, Lei A, Xu Y, Bi X, Huang X, Gao B, Wang L, Correia C, Chen M, Sun Q, Feng Y, Shen L, Wu H, Wang J, Shen X, Daley GQ, Li H, Zhang J LIN28 coordinately promotes nucleolar/ribosomal functions and represses the 2C-like transcriptional program in pluripotent stem cells. Protein Cell. 2022 Jul;13(7):490-512.
Junqueira C, Crespo Â, Ranjbar S, de Lacerda LB, Lewandrowski M, Ingber J, Parry B, Ravid S, Clark S, Schrimpf MR, Ho F, Beakes C, Margolin J, Russell N, Kays K, Boucau J, Das Adhikari U, Vora SM, Leger V, Gehrke L, Henderson LA, Janssen E, Kwon D, Sander C, Abraham J, Goldberg MB, Wu H, Mehta G, Bell S, Goldfeld AE, Filbin MR, Lieberman J. FcγR-mediated SARS-CoV-2 infection of monocytes activates inflammation. Nature. 2022 Jun;606(7914):576-584.
Meza-Sosa KF, Miao R, Navarro F, Zhang Z, Zhang Y, Hu JJ, Hartford CCR, Li XL, Pedraza-Alva G, Pérez-Martínez L, Lal A, Wu H, Lieberman J. SPARCLE, a p53-induced lncRNA, controls apoptosis after genotoxic stress by promoting PARP-1 cleavage. Mol Cell. 2022 Feb 17;82(4):785-802
Deng W, Bai Y, Deng F, Pan Y, Mei S, Zheng Z, Min R, Wu Z, Li W, Miao R, Zhang Z, Kupper TS, Lieberman J, Liu X. ( 2022 ) Streptococcal pyrogenic exotoxin B cleaves GSDMA and triggers pyroptosis. Nature. Feb 2;. doi: 10.1038/s41586-021-04384-4.
Gao W, Li Y, Liu X, Wang S, Mei P, Chen Z, Liu K, Li S, Xu XW, Gan J, Wu J, Ji C, Ding C, Liu X, Lai Y, He HH, Lieberman J, Wu H, Chen X, Li J. ( 2022 ) TRIM21 regulates pyroptotic cell death by promoting Gasdermin D oligomerization. Cell Death Differ. Feb;29(2):439-450.
Wang L, Crackower MA, Wu H. ( 2022 ) Advances toward structure-based drug discovery for inflammasome targets. J Exp Med. Jan 3;219(1). doi: 10.1084/jem.20211147.
Vora SM*, Lieberman J, Wu H† (2021). Inflammasome activation at the crux of severe COVID-19. Nat Rev Immunol. 21(11): 694-703.
Magupalli VG†, Fontana P, Wu H† (2021). Ragulator-Rag and ROS TORment gasdermin D pore formation. Trends Immunol. 42: 948-950.
Bittner ZA*, Liu X, Mateo Tortola M, Tapia-Abellán A, Shankar S, Andreeva L, Mangan M, Spalinger M, Kalbacher H, Düwell P, Lovotti M, Bosch K, Dickhöfer S, Marcu A, Stevanović S, Herster F, Cardona Gloria Y, Chang TH, Bork F, Greve CL, Löffler MW, Wolz OO, Schilling NA, Kümmerle-Deschner JB, Wagner S, Delor A, Grimbacher B, Hantschel O, Scharl M, Wu H†, Latz E†, Weber ANR†. (2021) . BTK operates a phospho-tyrosine switch to regulate NLRP3 inflammasome activity. J Exp Med. 218(11). doi: 10.1084/jem.20201656.
Fillmore N*, Bell S, Shen C, Nguyen V, La J, Dubreuil M, Strymish J, Brophy M, Mehta G, Wu H†, Lieberman J†, Do N†, Sander C†. (2021). Disulfiram use is associated with lower risk of COVID-19: (2021) A retrospective cohort study. PLoS One. 16(10): e0259061.
Tong AB, Burch JD, McKay D, Bustamante C, Crackower MA, Wu H. Could AlphaFold revolutionize chemical therapeutics?. Nat Struct Mol Biol. 2021 Oct;28(10):771-772. doi: 10.1038/s41594-021-00670-x. PubMed PMID: 34561631.
Gao W, Li Y, Liu X, Wang S, Mei P, Chen Z, Liu K, Li S, Xu XW, Gan J, Wu J, Ji C, Ding C, Liu X, Lai Y, He HH, Lieberman J, Wu H, Chen X, Li J. TRIM21 regulates pyroptotic cell death by promoting Gasdermin D oligomerization. Cell Death Differ. 2021 Sep 11;. doi: 10.1038/s41418-021-00867-z. [Epub ahead of print] PubMed PMID: 34511601.
Sun Z*, Yu H*, Zhao J*, Tan T, Pan H, Zhu Y, Chen L, Zhang C, Zhang L, Lei A, Xu Y, Bi X, Huang X, Gao B, Wang L, Correia C, Chen M, Sun Q, Feng Y, Shen L, Wu H, Wang J, Shen X, Daley GQ, Li H, Zhang J (2021). LIN28 coordinately promotes nucleolar/ribosomal functions and represses the 2C-like transcriptional program in pluripotent stem cells. Protein Cell . doi: 10.1007/s13238-021-00864-5.
Liu X*†, Xia S*, Zhang Z*, Wu H†, Lieberman J† (2021). Channeling inflammation: gasdermins in physiology and disease. Nat Rev Drug Discov. 20(5): 384-405.
Shen C*, Vohra M*, Zhang P, Mao X, Figley MD, Zhu J, Sasaki Y, Wu H†, DiAntonio A†, Milbrandt J†. (2021). Multiple domain interfaces mediate SARM1 autoinhibition. Proc Natl Acad Sci USA. 118(4): e2023151118.
Münzer P, Negro R, Fukui S, di Meglio L, Aymonnier K, Chu L, Cherpokova D, Gutch S, Sorvillo N, Shi L, Magupalli VG, Weber ANR, Scharf RE, Waterman CM, Wu H, Wagner DD†. (2021). NLRP3 Inflammasome Assembly in Neutrophils Is Supported by PAD4 and Promotes NETosis Under Sterile Conditions. Front Immunol 12: 683803
Wang L, Sharif H*, Vora SM*, Zheng Y, Wu H† (2021). Structures and functions of the inflammasome engine. J Allergy Clin Immunol 147: 2021-2029
Andreeva L, Wu H (2021). STING condensates on ER limit IFN response. Nat Cell Biol. 23: 299-300
Xia S*(2020). Biological mechanisms and therapeutic relevance of the gasdermin family. Mol Aspects Med. doi:10.1016/j.mam.2020.100890.
Hatcher JM, Yang G, Wang L, Ficarro SB, Buhrlage S, Wu H, Marto JA, Treon SP, Gray NS † (2020). Discovery of a Selective Covalent IRAK1 Inhibitor with Antiproliferative Activity in MYD88 Mutated B-Cell Lymphoma. ACS Med Chem Lett . doi: 10.1021/acsmedchemlett.0c00378
Wang L†, Wu D, Robinson CV, Wu H†, Fu TM†(2020). Structures of a Complete Human V-ATPase Reveal Mechanisms of Its Assembly. Mol Cell . doi: 10.1016/j.molcel.2020.09.029
Zhao L, Fu Q, Pan L, Piai A, Chou JJ †(2020). The diversity and similarity of transmembrane trimerization of TNF receptors. Front Cell Dev Biol. doi: 10.3389/fcell.2020.569684
Chen H*, Gu L*, Orellana EA, Wang Y, Guo J, Liu Q, Wang L, Shen Z, Wu H, Gregory RI, Xing Y, Shi Y† (2020). METTL4 is an snRNA m6Am methyltransferase that regulates RNA splicing. Cell Res doi: 10.1038/s41422-019-0270-4.
Xia S*, Hollingsworth LR 4th*, Wu H†(2019). Mechanism and Regulation of Gasdermin-Mediated Cell Death. Cold Spring Harb Perspect Biol. pii: a036400. doi: 10.1101/cshperspect.a036400.
Wu H*, Arnold E* (2019). Michael G. Rossmann (1930-2019) Nat Struct Mol Biol. (8):660-662.
Wang L, Wu H† (2019). Keeping the Death Protein in Check. Immunity. 51(1):1-2.
Scott DA*† , Hatcher JM, Liu H, Fu M, Du G, Fontán L, Us I, Casalena G, Qiao Q, Wu H, Melnick A, Gray NS (2019). Quinoline and thiazolopyridine allosteric inhibitors of MALT1. Bioorg Med Chem Lett. 29(14):1694-1698.
Arnold E†* , Wu H*, Johnson JE*. (2019). Michael G. Rossmann (1930-2019), pioneer in macromolecular and virus crystallography: scientist, mentor and friend. Acta Crystallogr D Struct Biol. 75(Pt 6):523-527.
Xia S*, Wang L*, Fu TM†, Wu H† (2019). Mechanism of TRPM2 channel gating revealed by cryo-EM. FEBS J. 286(17):3333-3339.
Xia S, Ruan J†, Wu H†(2019). Monitoring gasdermin pore formation in vitro. Methods Enzymol. 625:95-107
Shen C, Sharif H, Xia S, Wu H† (2019). Structural and mechanistic elucidation of inflammasome signaling by cryo-EM. Curr Opin Struct Biol. 58:18-25
Lee YR, Chen M, Lee JD*, Zhang J*, Lin SY*, Fu TM, Chen H, Ishikawa T, Chiang SY, Katon J, Zhang Y, Shulga YV, Bester AC, Fung J, Monteleone E, Wan L, Shen C, Hsu CH, Papa A, Clohessy JG, Teruya-Feldstein J, Jain S, Wu H, Matesic L, Chen RH, Wei W, Pandolfi PP† (2019). Reactivation of PTEN tumor suppressor for cancer treatment through inhibition of a MYC-WWP1 inhibitory pathway. Science. 364(6441).
Wang L*, Ferrao R*, Li Q, Hatcher JM, Choi HG, Buhrlage SJ, Gray NS, Wu H† (2019). Conformational flexibility and inhibitor binding to unphosphorylated interleukin-1 receptor-associated kinase 4 (IRAK4). The Journal of biological chemistry. 294(12):4511-4519.
Choi YJ, Kim S, Choi Y, Nielsen TB, Yan J, Lu A, Ruan J, Lee HR, Wu H, Spellberg B, Jung JU†(2019). SERPINB1-mediated checkpoint of inflammatory caspase activation. Nature immunology. 20(3):276-287.
Mompeán M*, Bozkurt G*, Wu H†.(2019). Mimicry by a viral RHIM. EMBO reports. 20(2).
Ru H, Zhang P, Wu H (2018). Structural gymnastics of RAG-mediated DNA cleavage in V(D)J recombination. Curr Opin Struct Biol. 53:178-186.
Orning P, Weng D1*, Starheim K*, Ratner D*, Best Z, Lee B, Brooks A, Xia S, Wu H, Kelliher MA, Berger SB, Gough PJ, Bertin J, Proulx MM, Goguen JD, Kayagaki N, Fitzgerald KA, Lien E†.(2018). Pathogen blockade of TAK1 triggers caspase-8–dependent cleavage of gasdermin D and cell death Science. 362(6418):1064-1069.
Luong P, Hedl M, Yan J, Zuo T, Fu TM, Jiang X, Thiagarajah JR, Hansen SH, Lesser CF, Wu H, Abraham C†, Lencer WI †(2018) INAVA-ARNO complexes bridge mucosal barrier function with inflammatory signaling. eLife. 2018; 7.
Fontán L*†, Qiao Q, Hatcher JM, Casalena G, Us I, Teater M, Durant M, Du G, Xia M, Bilchuk N, Chennamadhavuni S, Palladino G, Inghirami G, Philippar U, Wu H, Scott DA*, Gray NS, Melnick A*† (2018). Specific covalent inhibition of MALT1 paracaspase suppresses B cell lymphoma growth. The Journal of clinical investigation. 128(10):4397-4412.
Feltham R*, Jamal K*, Tenev T*, Liccardi G, Jaco I, Domingues CM, Morris O, John SW, Annibaldi A, Widya M, Kearney CJ, Clancy D, Elliott PR, Glatter T, Qiao Q, Thompson AJ, Nesvizhskii A, Schmidt A, Komander D, Wu H, Martin S, Meier P† (2018). Mind Bomb Refulates Cell Deth during TNF Signalling by Suppressing RIPK1’s Cytotoxic Potential. Cell Rep. 23(2):470-484.
Annibaldi A †, Wicky John S*, Vanden Berghe T*, Swatek KN*, Ruan J, Liccardi G, Bianchi K, Elliott PR, Choi SM, Van Coillie S, Bertin J, Wu H, Komander D, Vandenabeele P, Silke J, Meier P †(2018). Ubiquitin-Mediated Regulation of RIPK1 Kinase Activity Independent of IKK and MK2. Mol Cell. 69(4):566-580.
Xia S*, Fu TM*, Wu H† (2018). Inflammation NODs to Antagonists of RIP2-XIAP Interaction. Mol Cell. 69(4):535-536.
Fu TM*, Shen C*, Li Q, Zhang P, Wu H† (2018). Mechanism of ubiquitin transfer promoted by TRAF6. Proc Natl Acad Sci U S A. 115(8):1783-1788.
Zhang L, Wu H† (2018). Bad germs are trapped. Cell Res. 28(2):141-142
Evavold CL, Ruan J, Tan Y, Xia S, Wu H, Kagan JC† (2018). The Pore-Forming Protein Gasdermin D Regulates Interleukin-1 Secretion from Living Macrophages. Immunity. 48(1):35-44
Hauenstein AV, Xu G, Kabaleeswaran V, Wu H† (2017) .Evidence for M1-Linked Polyubiquitin-Mediated Conformational Change in NEMO. J Mol Biol429(24):3793-3800
Kleino A, Ramia NF, Bozkurt G, Shen Y, Nailwal H, Huang J, Napetschnig J, Gangloff M, Chan FK*, Wu H*†, Li J†, Silverman N*†.(2017). Peptidoglycan-Sensing Receptors Trigger the Formation of Functional Amyloids of the Adaptor Protein Imd to Initiate Drosophila NF-κB Signalin. Immunity 47(4):635-647
Wang H, Feng Z, Lu A, Jiang Y, Wu H, Xu B† (2017). Instant Hydrogelation Inspired by Inflammasomes. Angew Chem Int Ed Engl.56(26):7579-758
Wang L, Qiao Q, Wu H†(2017). Understanding CARD Tricks in Apoptosomes. Structure. 25(4)575-577
Yi L, Bozkurt G, Li Q, Lo S, Menon AK, Wu H† (2017). Disulfide Bond Formation and N-Glycosylation Modulate Protein-Protein Interactions in GPI-Transamidase (GPIT). Sci Rep. 8:45912
Lamour G, Nassar R*, Chan PH*, Bozkurt G, Li J, Bui JM, Yip CK, Mayor T, Li H, Wu H, Jorg A. Gsponer† (2017). Mapping the Broad Structural and Mechanical Properties of Amyloid Fibrils. Biophys. 112(4):584-594
Katja G. Weinacht*†, Louis-Marie Charbonnier*, Fayhan Alroqi, Ashley Plant, Qi Qiao, Hao Wu, Clement Ma, Troy R. Torgerson, Serogo D. Rosenzweig, Thomas A.Fleisher, Luigi D. Notarangelo, Imelda C. Hanson, Lisa R. Forbes, Talal A. Chatila (2017). Ruxolitinib reverses dysregulated T helper cell responses and controls autoimmunity caused by a novel signal transducer and activator of transcription 1 (STAT1) gain-of-function mutation. Journal of Allergy and Clinical Immunology. 139(5):1629