Cited by Preliminary exploration of the mechanisms underlying morphology engineering of Monascus purpureus during submerged fermentation via multi-omics approaches

Macrocyclic Peptide-Based Dual-Sensor Platform for Linkage-Specific Visualization of Ubiquitin Chain Assembling in Live Cells DOI

Han Gao,

Zhirong He,

Jian Chen

et al.

Analytical Chemistry, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 21, 2025

Intracellular monitoring of protein ubiquitination and differentiating polyubiquitin chain topology are crucial for understanding life processes drug discovery, which is challenged by the high complexity process a lack molecular tools. Herein, synthetic dual-sensor platform specific K48-linked ubiquitin oligomers was tailored in situ visualization assembling live biosystems. This achieved using macrocyclic peptides as recognition motifs tetraphenylethylene derivative an activatable reporter. The efficient cell penetration, tight binding, protection delivered "freeze-and-image" approach, allowing fluorescent readout linkage type elongation without perturbing physiological environment. Motivated these unique features, mapping K48-ubiquitination dynamics during degradation facilely achieved. Rapid, sensitive, intracellular assessment mechanism action potency dependence proteolysis-targeting chimeras (PROTACs) demonstrated, presenting sensors promising tools PROTAC development.

Language: Английский

Citations

A bacterial RING ubiquitin ligase triggering stepwise degradation of BRISC via TOLLIP-mediated selective autophagy manipulates host inflammatory response DOI

Xinming Pan, Yangyang Sun, Jianan Liu

et al.

Autophagy, Journal Year: 2025, Volume and Issue: unknown, P. 1 - 20

Published: Feb. 27, 2025

Numerous bacterial pathogens have evolved tactics to interfere with the host ubiquitination network evade clearance by innate immune system. Nevertheless, subtle antagonism between a ubiquitinase and deubiquitinase, through which they modify their respective targets within multifaceted network, has yet be characterized. BRCC3 isopeptidase complex (BRISC) is newly identified K63-specific deubiquitinase that plays crucial role in cellular signaling pathways such as inflammation. NleG, type III secretion system (T3SS) effector, contains conserved RING E3 ubiquitin ligase domain interacts machinery, along distinct substrate-recognition proteins. Here, one particular variant, NleG6, was mediating K27- K29-linked polyubiquitination at residues K89 K114 of ABRAXAS2/FAM175B, scaffolding protein BRISC complex, leading its degradation TOLLIP (toll interacting protein)-mediated selective autophagy. Further investigations elucidated ABRAXAS2 triggered subsequent adjacent BRCC3, turn, hindered TNIP1/ABIN1 degradation, ultimately inhibiting NFKB/NF-κB (nuclear factor kappa B)-mediated inflammatory responses. This chain events offers valuable insights into NFKB activation deubiquitinating BRISC, unveiling how bacteria manipulate regulation autophagy inhibit host's response thus dominate pathogen-host tug-of-war.Abbreviations: 3-MA: 3-methyladenine; A/E: attaching effacing; ATG7: related 7; BafA1: bafilomycin A1; BNIP3L/Nix: BCL2 3 like; BRISC: complex; Cas9: CRISPR-associated 9; co-IP: co-immunoprecipitation; CQ: chloroquine; CRISPR: clustered regulatory interspaced short palindromic repeat; DAPI: 4',6-diamidino2-phenylindole; DMSO: dimethyl sulfoxide; DUB: enzyme; E. coli: Escherichia coli; EHEC: enterohemorrhagic EPEC: enteropathogenic GFP: green fluorescent protein; LEE: locus enterocyte effacement; MAP1LC3B/LC3: microtubule associated 1 light beta; MG132: cbz-leu-leu-leucinal; MOI: multiplicity infection; NBR1: NBR1 cargo receptor; NC: negative control; NFKB/NF-κB: nuclear B; NH4Cl: ammonium chloride; OPTN: optineurin; SQSTM1/p62: sequestosome 1; sgRNAs: small guide RNAs; T3SS: system; TNF: tumor necrosis factor; TOLLIP: toll TRAF: TNF receptor TUBB: tubulin beta class I; WCL: whole cell lysate; WT: wide type.

Language: Английский

Citations

USP20 mitigates doxorubicin-induced cardiotoxicity by deubiquitinating and stabilizing HuR DOI

Zhouqing Huang, Yunxuan Chen, S S Wu

et al.

Research Square (Research Square), Journal Year: 2025, Volume and Issue: unknown

Published: April 16, 2025

Abstract Background The severe cardiotoxicity of doxorubicin (Dox) significantly restricts its clinical application. Deubiquitinating enzymes (DUBs) play pivotal roles in cardiac pathophysiology because their precise regulation protein function, localization and degradation.Objectives objective this study was to investigate the role molecular mechanism ubiquitin-specific peptidase 20 (USP20), a DUB, doxorubicin-induced cardiotoxicity.Methods Cardiomyocyte-specific USP20-knockout (USP20-CKO) mice were utilized assess USP20 cardiomyopathy (DIC). Coimmunoprecipitation (co-IP) combined with liquid chromatography‒mass spectrometry/mass spectrometry (LC‒MS/MS) analysis employed screen substrate USP20. Furthermore, mutant plasmids constructed elucidate underlying human antigen R (HuR) by Finally, an AAV9 vector used overexpress hearts cardiac-specific HuR-knockout interaction between HuR.Results results revealed decrease expression Dox-stimulated mouse cardiomyocytes. knockout resulted increased cardiomyocyte ferroptosis led DIC. Mechanistically, directly interacted HuR through protease structural domain. Deubiquitination at position 154 crucial for maintaining stability cleaving K48 ubiquitin chains inhibiting proteasomal degradation. Additionally, bound GPX4 mRNA suppress degradation, thereby mitigating contributing alleviating targeted overexpression via cardiomyocytes alleviated However, cardiomyocyte-specific knockout, no longer had anti-DIC effect, indicating that HuR, as downstream target USP20, plays irreplaceable DIC.Conclusions Our findings indicate enhances deubiquitination,

Language: Английский

Citations

Legionella effectors SidC/SdcA ubiquitinate multiple small GTPases and SNARE proteins to promote phagosomal maturation DOI

Kelong Ma,

Rundong Shu,

Hongtao Liu

et al.

Cellular and Molecular Life Sciences, Journal Year: 2024, Volume and Issue: 81(1)

Published: June 5, 2024

Protein ubiquitination is one of the most important posttranslational modifications (PTMs) in eukaryotes and involved regulation almost all cellular signaling pathways. The intracellular bacterial pathogen Legionella pneumophila translocates at least 26 effectors to hijack host via distinct mechanisms. Among these effectors, SidC/SdcA are novel E3 ubiquitin ligases with adoption a Cys-His-Asp catalytic triad. critical for recruitment endoplasmic reticulum (ER)-derived vesicles Legionella-containing vacuole (LCV). However, targets largely unknown, which restricts our understanding mechanisms used by vesicle trafficking pathway. Here, we demonstrated that multiple Rab small GTPases target soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNARE) proteins bona fide substrates SidC/SdcA. SidC/SdcA-mediated syntaxin 3 4 promotes their unconventional pairing vesicle-SNARE Sec22b, thereby contributing membrane fusion ER-derived phagosome. In addition, data reveal Rab7 its association LCV membrane. could impair binding downstream effector Rab-interacting lysosomal (RILP), partially explains why LCVs avoid lysosomes despite acquisition Rab7. Taken together, study reveals biological employed promote maturation LCVs.

Language: Английский

Citations

USP9X-mediated deubiquitination of Raptor contributes to autophagy impairment and memory deficits in P301S mice DOI

Siyi Zheng,

Jiahui Zhu, Cailin Wang

et al.

Cell Communication and Signaling, Journal Year: 2024, Volume and Issue: 22(1)

Published: Oct. 24, 2024

Language: Английский

Citations

Preliminary exploration of the mechanisms underlying morphology engineering of Monascus purpureus during submerged fermentation via multi-omics approaches DOI

Song Zhang,

Chenyu Zhang,

Xinyi Liu

et al.

LWT, Journal Year: 2024, Volume and Issue: unknown, P. 117108 - 117108

Published: Nov. 1, 2024

Language: Английский

Citations