iScience
·
2024
·
IF 4.6
Ziqi Wang,Tongtong Li,Dongkai Liu,Mengjing Li,Shangming Liu,Xiaochen Yu,Hanzhen Li,Hui Song,Wei Zhao,Zhaojian Liu,Xiangfeng Chen,Gang Lu,Zi-Jiang Chen,Tao Huang,Hongbin Liu
Spermiogenesis defines the final phase of male germ cell differentiation. While multiple deubiquitinating enzymes have been linked to spermiogenesis, the impacts of deubiquitination on spermiogenesis remain poorly characterized. Here, we investigated the function of UAF1 in mouse spermiogenesis. We selectively deleted Uaf1 in premeiotic germ cells using the Stra8-Cre knock-in mouse strain ( Uaf1 sKO), and found that Uaf1 is essential for spermiogenesis and male fertility. Further, UAF1 interacts and colocalizes with USP1 in the testes. Conditional knockout of Uaf1 in testes results in disturbed protein levels and localization of USP1, suggesting that UAF1 regulates spermiogenesis through the function of the deubiquitinating enzyme USP1. Using tandem mass tag-based proteomics, we identified that conditional knockout of Uaf1 in the testes results in reduced levels of proteins that are essential for spermiogenesis. Thus, we conclude that the UAF1/USP1 deubiquitinase complex is essential for normal spermiogenesis by regulating the levels of spermiogenesis-related proteins.
研究领域:
细胞生物学
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生殖医学
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分子生物学
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DOI
PubMed
高影响因子
Journal of Orthopaedic Surgery and Research
·
2023
·Vol. 18
·期 1
·1-12
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IF 2.6
Huang Jun,Zhou Hongxiang,He Liang,Zhong Lin,Zhou Ding,Yin Zongsheng
Background Nonunion is a failure of fracture healing and a major complication after fractures. Ubiquitin-specific protease 1 (USP1) is a deubiquitinase that involved in cell differentiation and cell response to DNA damage. Herein we investigated the expression, function and mechanism of USP1 in nonunion. Methods and results Clinical samples were used to detect the USP1 expression in nonunion. ML323 was selected to inhibit USP1 expression throughout the study. Rat models and mouse embryonic osteoblasts cells (MC3T3-E1) were used to investigate the effects of USP1 inhibition on fracture healing and osteogenesis in vivo and in vitro, respectively. Histological changes were examined by micro-computerized tomography (Micro-CT), hematoxylin & eosin (H&E) staining and Masson staining. Alkaline phosphatase (ALP) activity detection and alizarin red staining were used for osteogenic differentiation observation. The expression of related factors was detected by quantitative real-time PCR, western blot or immunohistochemistry (IHC). It was shown that USP1 was highly expressed in nonunion patients and nonunion rats. USP1 inhibition by ML323 promoted fracture healing in nonunion rats and facilitated the expression of osteogenesis-related factors and the signaling of PI3K/Akt pathway. In addition, USP1 inhibition accelerated osteogenic differentiation and promoting PI3K/Akt signaling in MC3T3-E1 cells. Conclusions USP1 inhibition plays a promotive role in coordinating osteogenic differentiation and fracture healing during nonunion. PI3K/Akt may be the downstream pathway of USP1.
研究领域:
骨科
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分子生物学
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细胞生物学
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DOI
PubMed
高影响因子
Microbiology Spectrum
·
2023
·Vol. 11
·期 3
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IF 3.7
Jinchao Xing,Chen Hu,Siqi Che,Yixin Lan,Lihong Huang,Lele Liu,Youqin Yin,Huanan Li,Ming Liao,Wenbao Qi
Japanese encephalitis virus (JEV) is a typical mosquito-borne flavivirus that can cause central nervous system diseases in humans and animals. Host factors attempt to limit virus replication when the viruses invade the host by using various strategies for replication. It is essential to clarify the host factors that affect the life cycle of JEV and explore its underlying mechanism. Here, we found that USP1-associated factor 1 (UAF1; also known as WD repeat-containing protein 48) modulated JEV replication. We found that JEV propagation significantly increased in UAF1-depleted Huh7 cells. Moreover, we found that knockdown of UAF1 activated cell autophagic flux in further functional analysis. Subsequently, we demonstrated that autophagy can be induced by JEV, which promotes viral replication by inhibiting interferon-stimulated gene (ISG) expression in Huh7 cells. The knockdown of UAF1 reduced ISG expression during JEV infection. To explore the possible roles of autophagy in UAF1-mediated inhibition of JEV propagation, we knocked out ATG7 to generate autophagy-deficient cells and found that depletion of UAF1 failed to promote JEV replication in ATG7 knockout cells. Moreover, in ATG7-deficient Huh7 cells, interference with UAF1 expression did not lead to the induction of autophagy. Taken together, these findings indicate that UAF1 is a critical regulator of autophagy and reveal a mechanism by which UAF1 knockdown activates autophagy to promote JEV replication.IMPORTANCE Host factors play an essential role in virus replication and pathogenesis. Although UAF1 is well known to form complexes with ubiquitin-specific proteases, little is known about the function of the UAF1 protein itself. In this study, we confirmed that UAF1 is involved in JEV replication. Notably, we discovered a novel function for UAF1 in regulating autophagy. Furthermore, we demonstrated that UAF1 modulated JEV replication through its autophagy regulation. This study is the first description of the novel function of UAF1 in regulating autophagy, and it clarifies the underlying mechanism of the antiviral effect of UAF1 against JEV. These results provide a new mechanistic insight into the functional annotation of UAF1 and provide a potential target for increasing virus production during vaccine production.
研究领域:
病毒学
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分子生物学
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细胞生物学
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传染病学
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PubMed
高影响因子
Translational Research
·
2024
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IF 6.4
Aoran Huang,Jianyun Rao,Xin Feng,Xingru Li,Tianhua Xu,Li Yao
Vascular calcification (VC) poses significant challenges in cardiovascular health. This study employs single-cell transcriptome sequencing to dissect cellular dynamics in this process. We identify distinct cell subgroups, notably in vascular smooth muscle cells (VSMCs), and observe differences between calcified atherosclerotic cores and adjacent regions. Further exploration reveals ID3 as a key gene regulating VSMC function. In vitro experiments demonstrate ID3′s interaction with USP1 and E12, modulating cell proliferation and osteogenic differentiation. Animal models confirm the critical role of the USP1/ID3/E12/P21 axis in VC. This study sheds light on a novel regulatory mechanism, offering potential therapeutic targets. 1. This study uses single-cell transcriptome to investigate vascular calcification in depth. 2. This study successfully identifies cellular heterogeneity related to vascular calcification. 3. This study reveals the crucial role of ID3 in vascular smooth muscle cell osteogenic differentiation. 4. In vivo and in vitro experiments of this study demonstrate the regulatory role of the USP1/ID3/E12/P21 axis in calcification. 5. This study provides potential molecular targets for the treatment of calcification-related diseases.
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高影响因子
INFLAMMATION
·
2026
·
IF 5.4
Wang Dongyan,Shi Yihai,Lu Jie,Bao Wen,Wang Nan,Shen Qiuteng,Lin Zhenyu,Qian Lijuan,Zhu Chang
Chronic pancreatitis (CP) is characterized by chronic pancreatic inflammation and progressive fibrosis. This study investigated the role of ubiquitin-specific peptidase 1 (USP1), a protein-stabilizing deubiquitinase, in CP. C57BL/6J mice were given 8-week repetitive intraperitoneal cerulein (50 µg/kg) injections to establish the CP model. USP1 expression in CP mouse pancreatic tissues was 3-fold higher than in controls. Lentivirus-mediated Usp1 knockdown reduced pancreatic inflammatory cell infiltration, trypsin activity, and proinflammatory cytokine levels. Additionally, Usp1 knockdown inhibits collagen deposition and fibrosis of the pancreas. In vitro, USP1 was upregulated in activated pancreatic stellate cells (PSCs) induced by TGF-β1 (5 ng/mL). USP1 knockdown decreased the expressions of collagen type I alpha 1 chain (COL1A1), COL1A2, fibronectin, and α-smooth muscle actin (α-SMA), suppressing PSC activation and extracellular matrix production. Immunoprecipitation-liquid chromatography/mass spectrometry (IP-LC/MS) and label-free proteomics identified integrin subunit beta 5 (ITGB5) as a potential target protein of USP1. USP1 promoted ITGB5 deubiquitination/stabilization, while USP1 knockdown reduced ITGB5 expression. Rescue experiments showed ITGB5 overexpression reversed the inhibitory effect of USP1 knockdown on PSC activation. Moreover, ITGB5 knockdown inhibited PSC activation via suppressing PI3K/AKT pathway. In conclusion, USP1 knockdown inhibits PSC activation by suppressing the ITGB5-PI3K-AKT axis, thereby alleviating the pathological progression of CP.
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高影响因子
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