MLKL-USP7-UBA52 signaling is indispensable for autophagy in brain through maintaining ubiquitin homeostasis

Zhang, Zhigang1,2; Chen, Shuai1,2,3; Jun, Shirui1,2; Xu, Xirong1,3; Hong, Yuchuan1,3; Yang, Xifei4; Zou, Liangyu5; Song, You-Qiang6; Chen, Yu1,2,3,7,8; Tu, Jie1,2,3,7,9

2024-09-01

10.1080/15548627.2024.2395727

AUTOPHAGY   影响因子和分区

1554-8627

1554-8635

Individuals with genetic elimination of MLKL (mixed lineage kinase domain like pseudokinase) exhibit an increased susceptibility to neurodegenerative diseases like Alzheimer disease (AD). However, the mechanism is not yet fully understood. Here, we observed significant compromise in macroautophagy/autophagy in the brains of mlkl knockout (KO) mice, as evidenced by the downregulation of BECN1/Beclin1 and ULK1 (unc-51 like autophagy activating kinase 1). We identified UBA52 (ubiquitin A-52 residue ribosomal protein fusion product 1) as the binding partner of MLKL under physiological conditions. Loss of Mlkl induced a decrease in ubiquitin levels by preventing UBA52 cleavage. Furthermore, we demonstrated that the deubiquitinase (DUB) USP7 (ubiquitin specific peptidase 7) mediates the processing of UBA52, which is regulated by MLKL. Moreover, our results indicated that the reduction of BECN1 and ULK1 upon Mlkl loss is attributed to a decrease in their lysine 63 (K63)-linked polyubiquitination. Additionally, single-nucleus RNA sequencing revealed that the loss of Mlkl resulted in the disruption of multiple neurodegenerative disease-related pathways, including those associated with AD. These results were consistent with the observation of cognitive impairment in mlkl KO mice and exacerbation of AD pathologies in an AD mouse model with mlkl deletion. Taken together, our findings demonstrate that MLKL-USP7-UBA52 signaling is required for autophagy in brain through maintaining ubiquitin homeostasis, and highlight the contribution of Mlkl loss-induced ubiquitin deficits to the development of neurodegeneration. Thus, the maintenance of adequate levels of ubiquitin may provide a novel perspective to protect individuals from multiple neurodegenerative diseases through regulating autophagy.Abbreviations: 4HB: four-helix bundle; AAV: adeno-associated virus; AD: Alzheimer disease; AIF1: allograft inflammatory factor 1; APOE: apolipoprotein E; APP: amyloid beta precursor protein; A beta: amyloid beta; BECN1: beclin 1; co-IP: co-immunoprecipitation; DEGs: differentially expressed genes; DLG4: discs large MAGUK scaffold protein 4; DUB: deubiquitinase; EBSS: Earle's balanced salt solution; GFAP: glial fibrillary acidic protein; HRP: horseradish peroxidase; IL1B: interleukin 1 beta; IL6: interleukin 6; IPed: immunoprecipitated; KEGG: Kyoto Encyclopedia of Genes and Genomes; KO: knockout; MAP1LC3/LC3: microtubule associated protein 1 light chain 3; MLKL: mixed lineage kinase domain like pseudokinase; NSA: necrosulfonamide; OPCs: oligodendrocyte precursor cells; PFA: paraformaldehyde; PsKD: pseudo-kinase domain; SYP: synaptophysin; UB: ubiquitin; UBA52: ubiquitin A-52 residue ribosomal protein fusion product 1; UCHL3: ubiquitin C-terminal hydrolase L3; ULK1: unc-51 like autophagy activating kinase 1; UMAP: uniform manifold approximation and projection; UPS: ubiquitin-proteasome system; USP7: ubiquitin specific peptidase 7; USP9X: ubiquitin specific peptidase 9 X-linked.

Alzheimer disease autophagy BECN1 cognitive impairment neurodegeneration ULK1

SCI

英语

其他

Key-Area Research and Development Program of Guangdong Province[2023B0303040004] ; National Natural Science Foundation of China [NSFC][32371070] ; NSFC-RGC Joint Research Scheme[32061160472] ; Guangdong Provincial Fund for Basic and Applied Basic Research[2019B1515130004] ; Hong Kong Innovation and Technology Fund[MRP/056/21] ; Science and Technology Program of Guangzhou[202007030001] ; Shenzhen Key Laboratory of Neuroimmunomodulation for Neurological Diseases[ZDSYS20220304163558001] ; Key Basic Research Program of Shenzhen Science and Technology Innovation Commission["JCYJ20210324115811031","JCYJ20220818101618040","JCYJ20220818101615033","JCYJ20210324101813035","JCYJ20200109150717745","JCYJ20200109144418639"]

Cell Biology

Cell Biology

WOS:001316004800001

TAYLOR & FRANCIS INC

Web of Science

1.Chinese Acad Sci, Shenzhen Hong Kong Inst Brain Sci, Shenzhen Inst Adv Technol, Shenzhen Key Lab Neuroimmunomodulat Neurol Dis, Shenzhen, Peoples R China
2.Shenzhen Univ Adv Technol, Fac Life & Hlth Sci, Shenzhen, Guangdong, Peoples R China
3.Univ Chinese Acad Sci, Beijing, Peoples R China
4.Shenzhen Ctr Dis Control & Prevent, Key Lab Modern Toxicol Shenzhen, Shenzhen, Peoples R China
5.Southern Univ Sci & Technol, Jinan Univ, Shenzhen Peoples Hosp, Affiliated Hosp 1,Clin Coll 2,Dept Neurol, Shenzhen, Peoples R China
6.Univ Hong Kong, Sch Biomed Sci, 21 Sassoon Rd, Hong Kong, Peoples R China
7.Chinese Acad Sci, Shenzhen Inst Adv Technol, CAS Key Lab Brain Connectome & Manipulat, Shenzhen, Peoples R China
8.Chinese Acad Sci, SIAT HKUST Joint Lab Brain Sci, Shenzhen, Peoples R China
9.Chinese Acad Sci, Shenzhen Inst Adv Technol, Guangdong Prov Key Lab Brain Connectome & Behav, Shenzhen, Guangdong, Peoples R China

Zhang, Zhigang,Chen, Shuai,Jun, Shirui,et al. MLKL-USP7-UBA52 signaling is indispensable for autophagy in brain through maintaining ubiquitin homeostasis[J]. AUTOPHAGY,2024.

Zhang, Zhigang.,Chen, Shuai.,Jun, Shirui.,Xu, Xirong.,Hong, Yuchuan.,...&Tu, Jie.(2024).MLKL-USP7-UBA52 signaling is indispensable for autophagy in brain through maintaining ubiquitin homeostasis.AUTOPHAGY.

Zhang, Zhigang,et al."MLKL-USP7-UBA52 signaling is indispensable for autophagy in brain through maintaining ubiquitin homeostasis".AUTOPHAGY (2024).