Biochimica et biophysica acta. Molecular cell research最新文献_第3页

Mutant-specific dysfunction of RHOBTB2 impairs mitochondrial function and Na+/K+-ATPase levels in a cell model 在细胞模型中，RHOBTB2突变特异性功能障碍损害线粒体功能和Na+/K+- atp酶水平

IF 3.7 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2025-12-31 DOI: 10.1016/j.bbamcr.2025.120103

Sachiko Miyamoto , Shun-ichi Yamashita , Hazrat Belal , Mitsuhiro Kato , Tomotake Kanki , Hirotomo Saitsu

RHOBTB2 is an atypical Rho GTPase implicated in developmental and epileptic encephalopathy, yet its pathogenic mechanisms remain poorly understood. In this study, we established a cell model in which RHOBTB2 expression was induced by a doxycycline-inducible system to investigate the functional consequences of disease-associated RHOBTB2 mutants. Protein expression and localization analyses revealed mutant-specific behaviors: GTPase-domain mutants such as D92H and W217C showed no significant difference in RHOBTB2 protein levels compared with WT, while hotspot mutants (R461H, R485C, R489Q) exhibited increased RHOBTB2 protein levels with nuclear and mitochondrial accumulation. RNA-seq analysis revealed that cells expressing each mutant showed distinct transcriptomic changes. Notably, induction of the R489Q mutant caused a robust downregulation of ion channel-related genes, supporting its potential role in disrupting neuronal excitability. Furthermore, expression of mutant RHOBTB2 led to a reduction in Na⁺/K⁺-ATPase protein levels via a lysosome-dependent degradation pathway. This effect was particularly prominent in GTPase-domain mutants (D92H and W217C), suggesting a mechanistic link between mutant RHOBTB2 and impaired ion homeostasis. Moreover, the R489Q and W217C mutants impaired mitochondrial respiration, whereas other mutants did not show detectable mitochondrial dysfunction. Importantly, the Y284D cancer-associated mutant did not share these phenotypes, highlighting the diversity of functional outcomes across different mutation sites. The observed dysregulation of ion transport pathways and mitochondrial impairment in some mutants may represent key mechanisms underlying seizure susceptibility and other neurological manifestations in RHOBTB2-associated encephalopathies.

RHOBTB2是一种与发育性和癫痫性脑病有关的非典型Rho GTPase，但其致病机制尚不清楚。在这项研究中，我们建立了一个由强力霉素诱导系统诱导RHOBTB2表达的细胞模型，以研究疾病相关的RHOBTB2突变体的功能后果。蛋白表达和定位分析揭示了突变的特异性行为：gtpase结构域突变体如D92H和W217C与WT相比RHOBTB2蛋白水平无显著差异，而热点突变体（R461H、R485C、R489Q） RHOBTB2蛋白水平升高，并有核和线粒体积累。RNA-seq分析显示，表达每种突变的细胞表现出明显的转录组变化。值得注意的是，R489Q突变体的诱导导致离子通道相关基因的强烈下调，支持其在破坏神经元兴奋性中的潜在作用。此外，RHOBTB2突变体的表达通过溶酶体依赖的降解途径导致Na+/K+- atp酶蛋白水平的降低。这种效应在gtpase结构域突变体（D92H和W217C）中尤为突出，表明突变体RHOBTB2与离子稳态受损之间存在机制联系。此外，R489Q和W217C突变体损伤了线粒体呼吸，而其他突变体没有表现出可检测到的线粒体功能障碍。重要的是，Y284D癌症相关突变体没有共享这些表型，突出了不同突变位点的功能结果的多样性。在一些突变体中观察到的离子转运通路失调和线粒体损伤可能是rhobtb2相关脑病中癫痫易感性和其他神经学表现的关键机制。

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引用次数: 0

Prevotella copri leads to colonic barrier dysfunction via the succinate receptor 1-FoxM1-IL-6 axis copri普雷沃氏菌通过琥珀酸受体1-FoxM1-IL-6轴导致结肠屏障功能障碍。

IF 3.7 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2025-12-30 DOI: 10.1016/j.bbamcr.2025.120099

Rui Li , Shuang Liu , Haiyan Zhang , Xinglin Huang , Wenchu Qian , Daobin Han , Fang Wang , Zhonghui Feng , Tongtong Zhang , Hai Lin , Haifang Li

Prevotella copri, a key commensal bacterium in the human gut microbiome, exhibits both positive and negative abundance correlations with various disorders. Although multiple studies have suggested an association between P. copri and intestinal pathologies, the mechanistic basis remains elusive. In this study, we examined P. copri's effects on colonic physiology in healthy mice, revealing its capacity to compromise intestinal barrier integrity. This was demonstrated through downregulation of colonic tight junction proteins (Occludin and ZO-1) and elevated serum levels of gut permeability markers (LPS and D-LA). We identified succinate as the primary microbial metabolite mediating P. copri's barrier-disrupting effects, with succinate receptor 1 (SUCNR1) being essential for this pathological process. Notably, both P. copri colonization and succinate administration activated the IL-6-STAT3 signaling pathway, leading to transcriptional suppression of the tight junction-related gene, Occludin. Through mechanistic studies, we identified Forkhead box M1 (FoxM1) as a crucial transcription factor regulating P. copri- or succinate-induced Il-6 expression. Clinical relevance was established through human cohort analyses showing significant positive correlations between fecal succinate levels and plasma markers of gut permeability. These findings elucidate a novel mechanistic pathway through which P. copri impairs colonic function, suggesting therapeutic potential in colitis through strategies targeting either P. copri abundance or microbial succinate production.

copri普雷沃特菌是人类肠道微生物群中的一种重要的共生细菌，其丰度与多种疾病呈正相关和负相关。尽管多项研究表明copri与肠道病变之间存在关联，但其机制基础仍难以捉摸。在这项研究中，我们检测了copri对健康小鼠结肠生理的影响，揭示了其损害肠道屏障完整性的能力。这可以通过结肠紧密连接蛋白（Occludin和ZO-1）的下调和肠道通透性标志物（LPS和D-LA）的血清水平升高来证明。我们发现琥珀酸盐是介导copri屏障破坏作用的主要微生物代谢物，琥珀酸受体1 （SUCNR1）在这一病理过程中至关重要。值得注意的是，copri定植和琥珀酸盐给药都激活了IL-6-STAT3信号通路，导致紧密连接相关基因Occludin的转录抑制。通过机制研究，我们确定叉头盒M1 （FoxM1）是调节P. copri或琥珀酸诱导的Il-6表达的关键转录因子。通过人类队列分析，建立了临床相关性，显示粪便琥珀酸盐水平与肠道通透性血浆标志物之间存在显著的正相关。这些发现阐明了copri损害结肠功能的一种新的机制途径，提示通过针对copri丰度或微生物琥珀酸盐产生的策略治疗结肠炎的潜力。

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引用次数: 0

NSUN2 activated by KLF6 can mediate m5C modification of NCOA4 to enhance ferroptosis of OGD/R-induced SH-SY5Y cells in ischemic stroke KLF6激活的NSUN2可介导NCOA4的m5C修饰，增强OGD/ r诱导的缺血性卒中SH-SY5Y细胞的铁凋亡。

IF 3.7 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2025-12-30 DOI: 10.1016/j.bbamcr.2025.120102

Genshan Gao , Yue Xu , Yao Liu, Shuqing Meng, Nannuan Liu

Ischemic stroke (IS) constitutes the majority of stroke cases. Ferroptosis, a non-apoptotic form of programmed cell death, is an essential mechanism of IS. This study aimed to investigate a ferroptosis-related molecular mechanism in IS progression. IS model in vitro was induced by oxygen-glucose deprivation/reoxygenation (OGD/R) in SH-SY5Y cells. RT-qPCR and Western blotting were performed for detection of NOP2/Sun RNA methyltransferase family member 2 (NSUN2), nuclear receptor coactivator 4 (NCOA4), and Krueppel-like factor 6 (KLF6). Cell functions were assessed by MTT assay, flow cytometry, and enzyme-linked immunosorbent assay. Mitochondrial membrane potential and other ferroptosis indicators were examined using kits. Molecular binding was detected using methylated RNA immunoprecipitation (MeRIP), RIP, dual-luciferase reporter assay and chromatin immunoprecipitation (ChIP) assay. IS animal model was established by middle cerebral artery occlusion (MCAO). OGD/R-induced apoptosis, inflammation, and ferroptosis of SH-SY5Y cells were suppressed by NCOA4 knockdown. NSUN2 could promote NCOA4 mRNA stability by mediating 5-methylcytosine (m5C) methylation modification of NCOA4, and YBX1 served as a reader protein. NSUN2 contributed to OGD/R-induced SH-SY5Y cell injury via upregulating NCOA4. KLF6 acted as a transcription factor to activate NSUN2 transcription, and then facilitated nerve injury of OGD/R-induced SH-SY5Y cells. Animal assay showed that silencing NSUN2 inhibited infarct volume, tissue injury, neurological function, and neuroinflammation in MCAO rats. These current findings affirmed that KLF6-activated NSUN2 could contribute to ferroptosis of OGD/R-induced SH-SY5Y cells via inducing NCOA4 m5C modification, providing a novel insight into the mechanism of ferroptosis in IS.

缺血性中风（IS）占中风病例的大多数。铁凋亡是一种非凋亡形式的程序性细胞死亡，是is的重要机制。本研究旨在探讨IS进展中与铁衰相关的分子机制。采用氧-葡萄糖剥夺/再氧化（OGD/R）法诱导SH-SY5Y细胞体外IS模型。采用RT-qPCR和Western blotting检测NOP2/Sun RNA甲基转移酶家族成员2 （NSUN2）、核受体辅助激活因子4 （NCOA4）和krueppel样因子6 （KLF6）。采用MTT法、流式细胞术和酶联免疫吸附法评估细胞功能。采用试剂盒检测线粒体膜电位及其他铁下垂指标。采用甲基化RNA免疫沉淀法（MeRIP）、RIP、双荧光素酶报告基因法和染色质免疫沉淀法（ChIP）检测分子结合。采用大脑中动脉闭塞法（MCAO）建立IS动物模型。下调NCOA4可抑制OGD/ r诱导的SH-SY5Y细胞凋亡、炎症和铁下垂。NSUN2可通过介导NCOA4的5-甲基胞嘧啶（m5C）甲基化修饰，促进NCOA4 mRNA的稳定性，YBX1作为一个读取器蛋白。NSUN2通过上调NCOA4参与OGD/ r诱导的SH-SY5Y细胞损伤。KLF6作为转录因子激活NSUN2转录，进而促进OGD/ r诱导的SH-SY5Y细胞的神经损伤。动物实验表明，沉默NSUN2可抑制MCAO大鼠的梗死体积、组织损伤、神经功能和神经炎症。这些研究结果证实了klf6激活的NSUN2可以通过诱导NCOA4 m5C修饰促进OGD/ r诱导的SH-SY5Y细胞的铁死亡，为IS中铁死亡的机制提供了新的见解。

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引用次数: 0

The tumor suppressor role of mitochondrial E3 ubiquitin ligase MUL1 in osteosarcoma 线粒体E3泛素连接酶MUL1在骨肉瘤中的抑瘤作用。

IF 3.7 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2025-12-28 DOI: 10.1016/j.bbamcr.2025.120101

Jacopo Di Gregorio , Sara Terreri , Michela Rossi , Giulia Battafarano , Laura Di Giuseppe , Olivia Pagliarosi , Lucia Cilenti , Enrico Ricevuto , Antonis S. Zervos , Vincenzo Flati , Andrea Del Fattore

Osteosarcoma is a highly aggressive type of bone cancer with a high rate of metastasis. The molecular mechanisms underlying osteosarcoma metastasis are not yet completely understood, representing an ongoing challenge for therapy. A possible therapeutic target is the hypoxia-inducible factor HIF-1α which is upregulated in metastatic osteosarcoma. Indeed, HIF-1α promotes proliferation, resistance to apoptosis and metabolic reprogramming towards glycolysis, whereas its downregulation increases apoptosis. The molecular mechanism mediated by the mitochondrial E3 ubiquitin ligase MUL1 could be exploited to target HIF-1α since low MUL1 protein levels result in HIF-1α accumulation and activity even under normoxic conditions, while high levels of MUL1 promote HIF-1α degradation. Here, we show that MUL1 protein levels inversely correlate with the aggressiveness of osteosarcoma cell lines. Induction of MUL1 in aggressive cells reduces HIF-1α levels, paired with a decrease in proliferation, migration and glycolysis and increase in apoptosis, whereas MUL1 inactivation in low-aggressive cells has opposite results. Therefore, the modulation of MUL1 protein levels affects cell proliferation, migration, apoptosis, and metabolism. This is the first report that reveals a tumor suppressor role for MUL1 in osteosarcoma, and suggests MUL1 induction as a potential therapeutic strategy to reduce HIF-1α activity in the metastatic progression of this cancer.

骨肉瘤是一种高度侵袭性的骨癌，具有很高的转移率。骨肉瘤转移的分子机制尚不完全清楚，这对治疗来说是一个持续的挑战。一个可能的治疗靶点是缺氧诱导因子HIF-1α，它在转移性骨肉瘤中上调。事实上，HIF-1α促进细胞增殖、抗凋亡和糖酵解代谢重编程，而其下调则增加细胞凋亡。线粒体E3泛素连接酶MUL1介导的分子机制可以被利用来靶向HIF-1α，因为低MUL1蛋白水平即使在常温条件下也会导致HIF-1α积累和活性，而高水平的MUL1促进HIF-1α降解。在这里，我们发现MUL1蛋白水平与骨肉瘤细胞系的侵袭性呈负相关。在侵袭性细胞中，MUL1的诱导降低了HIF-1α水平，同时伴随着增殖、迁移和糖酵解的减少以及细胞凋亡的增加，而在低侵袭性细胞中，MUL1的失活则有相反的结果。因此，MUL1蛋白水平的调节影响细胞增殖、迁移、凋亡和代谢。这是首次报道MUL1在骨肉瘤中的抑瘤作用，并提示MUL1诱导作为一种潜在的治疗策略来降低这种癌症转移进展中的HIF-1α活性。

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引用次数: 0

Cationic liposome-based silencing of thioredoxin reductase-1 for sensitization of MDA-MB 231 cells to gamma radiation 基于阳离子脂质体的硫氧还蛋白还原酶-1沉默对MDA-MB 231细胞对伽马辐射的增敏作用。

IF 3.7 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2025-12-26 DOI: 10.1016/j.bbamcr.2025.120100

Sandeep B. Shelar , Manidipa Basu , P.A. Hassan , K.C. Barick

Thioredoxin reductase 1 (TrxRd1) is a widely conserved antioxidant enzyme that maintains cellular redox homeostasis by reducing oxidized thioredoxin 1 (Trx1). Overexpression of TrxRd1 has been directly linked to radioresistance in cancer cells. In this study, we examined the impact of cationic liposome-mediated knockdown of TrxRd1 on the radiosensitization of MDA-MB 231 breast cancer cells. Cationic liposomes were formulated using dihexadecyldimethylammonium bromide (DDAB) as a surfactant and soy lecithin as the lipid component, employing a supercooled micelle-based method. These liposomes were complexed with plasmid DNA encoding a short hairpin RNA targeting TrxRd1 (shTrxRd1) via electrostatic interactions. Lipoplex formation (DDAB-shTrxRd1) was confirmed by assessing changes in hydrodynamic size, surface charge, and isothermal titration calorimetry. Cellular uptake was demonstrated by cytosolic localization of propidium iodide (PI) and expression of green fluorescent protein (GFP), while reduced TrxRd1 mRNA levels confirmed gene silencing. TrxRd1 knockdown led to elevated reactive oxygen species (ROS) levels in gamma-irradiated cells. This was accompanied by decreased cell viability, reduced colony formation, and increased apoptosis, indicating enhanced radiosensitization of MDA-MB 231 cells following TrxRd1 suppression and gamma radiation exposure. In summary, targeting TxrRd1 by cationic liposome gene silencing provide improved radiosensitization of breast cancer cells through augmenting ROS accumulation.

硫氧还蛋白还原酶1 （TrxRd1）是一种广泛保守的抗氧化酶，通过还原氧化的硫氧还蛋白1 （Trx1）来维持细胞氧化还原稳态。TrxRd1的过表达与癌细胞的放射耐药直接相关。在这项研究中，我们检测了阳离子脂质体介导的TrxRd1下调对MDA-MB 231乳腺癌细胞放射增敏的影响。以二十六烷基二甲基溴化铵（DDAB）为表面活性剂，大豆卵磷脂为脂质组分，采用过冷胶束法制备阳离子脂质体。这些脂质体通过静电相互作用与编码短发夹RNA靶向TrxRd1 （shTrxRd1）的质粒DNA复合物。脂质体形成（DDAB-shTrxRd1）通过评估流体动力学大小、表面电荷和等温滴定量热法的变化得到证实。通过碘化丙啶（PI）的胞质定位和绿色荧光蛋白（GFP）的表达证实了细胞摄取，而TrxRd1 mRNA水平的降低证实了基因沉默。TrxRd1敲低导致γ辐照细胞中活性氧（ROS）水平升高。这伴随着细胞活力下降、集落形成减少和凋亡增加，表明TrxRd1抑制和γ辐射暴露后MDA-MB 231细胞的放射增敏性增强。综上所述，通过阳离子脂质体基因沉默靶向TxrRd1可通过增加ROS积累改善乳腺癌细胞的放射增敏。

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引用次数: 0

Ephrin-A1/EphA2 modulates vascular smooth muscle cell proliferation in vitro, while VSMC-specific ephrin-A1 deletion is not sufficient to alter arteriosclerosis in vivo Ephrin-A1/EphA2在体外调节血管平滑肌细胞增殖，而vsmc特异性的Ephrin-A1缺失不足以改变体内动脉硬化。

IF 3.7 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2025-12-22 DOI: 10.1016/j.bbamcr.2025.120098

Antje Augstein , Sebastian Porth , Johannes Mierke , Janet Friedrich , Stefanie Jellinghaus , Axel Linke , Volker Adams , David M. Poitz

Atherosclerosis, the leading cause of heart attack and stroke, involves plaque formation driven by various cell types, including endothelial cells, immune cells and vascular smooth muscle cells (VSMCs). VSMCs undergo a phenotypic switch from a contractile to a synthetic state, contributing to disease progression. The Eph/ephrin signaling pathway, particularly ephrin-A1 and its receptor EphA2, has been implicated in this phenotypic modulation. Its role in atherosclerosis was explored using in vitro and in vivo models.

In plaque-containing arteries, both ephrin-A1 and EphA2 were upregulated compared to plaque-free regions. The phenotypic transition of VSMCs from a contractile to a synthetic state is associated with reduced ephrin-A1 levels, elevated EphA2 expression, and increased cellular proliferation. A consistent expression pattern with low ephrin-A1 and high EphA2 was observed in proliferating cells. Additionally, under pro-proliferative conditions, the non-canonical phosphorylation site S897/898 of EphA2 is phosphorylated in a MEK/ERK/RSK-dependent manner, while the canonical site Y588/589 undergoes autophosphorylation at higher ephrin-A1 levels. These in vitro findings suggest an anti-proliferative, potentially anti-atherogenic role for ephrin-A1 in VSMCs. This hypothesis was further examined in ApoE-KO mice using a conditional VSMC-specific ephrin-A1 knockout. Surprisingly, neither high-fat diet-induced atherosclerosis nor wire injury-induced stenosis differed between ephrin-A1-deficient and wild-type mice, implying functional compensation by other ephrin ligands.

The present findings highlight significant roles for the Eph/ephrin system in VSMC biology and plaque development. However, its effects appear to be multimodal, influenced by the interplay between various cell types and distinct members of the Eph/ephrin family.

动脉粥样硬化是心脏病发作和中风的主要原因，涉及由多种细胞类型驱动的斑块形成，包括内皮细胞、免疫细胞和血管平滑肌细胞（VSMCs）。VSMCs经历从收缩状态到合成状态的表型转换，有助于疾病进展。Eph/ephrin信号通路，特别是ephrin- a1及其受体EphA2，参与了这种表型调节。通过体外和体内模型探讨其在动脉粥样硬化中的作用。在含斑块动脉中，与无斑块区相比，ephrin-A1和EphA2均上调。VSMCs从收缩状态到合成状态的表型转变与ephrin-A1水平降低、EphA2表达升高和细胞增殖增加有关。增殖细胞中ephrin-A1低表达，EphA2高表达。此外，在促增殖条件下，EphA2的非典型磷酸化位点S897/898以MEK/ERK/ rsk依赖的方式磷酸化，而典型位点Y588/589在较高的ephrin-A1水平下进行自磷酸化。这些体外研究结果表明，在VSMCs中，ephrin-A1具有抗增殖、潜在的抗动脉粥样硬化作用。这一假设在ApoE-KO小鼠中进一步验证，使用条件vsmc特异性ephrin-A1敲除。令人惊讶的是，高脂肪饮食诱导的动脉粥样硬化和导线损伤诱导的狭窄在ephrin- a1缺陷小鼠和野生型小鼠之间都没有差异，这意味着其他ephrin配体的功能补偿。目前的研究结果强调了Eph/ephrin系统在VSMC生物学和斑块发育中的重要作用。然而，它的作用似乎是多模式的，受各种细胞类型和Eph/ephrin家族不同成员之间相互作用的影响。

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引用次数: 0

RelK, a YoeB-like toxin from Mycobacterium tuberculosis displays ribosome independent endonucleolytic activity 来自结核分枝杆菌的一种类似yoeb的毒素RelK显示出核糖体独立的核内溶活性。

IF 3.7 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2025-12-19 DOI: 10.1016/j.bbamcr.2025.120097

Shafinaz Rahman Sarah , Nimisha Sinha , Harsh A. Gandhi , Jaydeep Bhattacharya , Vandana Malhotra

Type II toxin-antitoxin (TA) modules are paired genetic elements encoding a toxin protein and an associated antitoxin that neutralizes the toxin under favourable growth conditions. Stress-induced degradation of the antitoxin renders the toxin free to disrupt essential cellular processes leading to growth arrest or cell death. Mycobacterium tuberculosis (M. tb) is known to harbor 60+ such modules that contribute to its pathogenicity and persistence. The RelK toxin of the RelJK cassette has generated significant interest given its expression during infection and regulation by post-translational modification; however, its functional activity remains uncharacterized. Using a cell free transcription-translation system we show that both, unphosphorylated and phosphorylated RelK toxin inhibit in vitro protein synthesis implicating a role in translational control, one that is enhanced by S/T phosphorylation. A key finding of this study is the ribosome-independent nuclease activity of RelK. We discovered that in addition to the in vitro RNase activity, RelK also cleaves double-stranded DNA in a dose-dependent manner, and is inactivated by the RelJ antitoxin. Further characterization established RelK as a nickase whose activity is modulated by divalent cations, and is independent of the substrate topology, although supercoiled DNA substrates are preferred. Molecular docking of RelK revealed multiple contacts with the phosphate backbone and bases of the dsDNA facilitating binding and protein orientation on the major groove region. Furthermore, substituting His84 with glutamine in RelK not only abolished its catalytic activity but also obliterated cytotoxicity. The data highlights the diversity in substrates and catalytic activities of M. tb Type II toxins.

II型毒素-抗毒素（TA）模块是成对的遗传元件，编码毒素蛋白和相关的抗毒素，在有利的生长条件下中和毒素。应激诱导的抗毒素降解使毒素自由地破坏基本的细胞过程，导致生长停滞或细胞死亡。已知结核分枝杆菌（M. tb）含有60多个这样的模块，这些模块有助于其致病性和持久性。RelJK卡带的RelK毒素在感染期间的表达和翻译后修饰的调节引起了极大的兴趣；然而，它的功能活动仍然是未知的。利用细胞自由转录-翻译系统，我们发现未磷酸化和磷酸化的RelK毒素都能抑制体外蛋白质合成，这暗示了翻译控制的作用，而S/T磷酸化则能增强这种作用。这项研究的一个关键发现是RelK的核糖体非依赖性核酸酶活性。我们发现，除了体外RNase活性外，RelK还以剂量依赖的方式切割双链DNA，并被RelJ抗毒素灭活。进一步的表征表明，RelK是一种酶，其活性由二价阳离子调节，并且与底物拓扑结构无关，尽管超螺旋DNA底物是首选。RelK的分子对接揭示了它与dsDNA的磷酸主链和碱基的多重接触，促进了主要凹槽区域的结合和蛋白质取向。此外，用谷氨酰胺取代RelK中的His84不仅可以消除其催化活性，还可以消除其细胞毒性。数据强调了结核分枝杆菌II型毒素底物和催化活性的多样性。

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引用次数: 0

Sustained Galiellalactone treatment breaks the metastatic resilience of BRAF-inhibitor-resistant melanoma cells 持续的盖ellalactone治疗打破braf抑制剂抵抗黑色素瘤细胞的转移弹性

IF 3.7 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2025-12-10 DOI: 10.1016/j.bbamcr.2025.120096

Sujoy Bhowmik, Saumya Ranjan Badajena , Satyajit Laxman Murkute , Dumarapu Bhagyalaxmi, Purusottam Mohapatra

STAT3 hyperactivation in metastatic and therapy-resistant melanoma drives tumorigenesis and immunotherapy resistance by promoting tumor-stromal interactions, making its inhibition a crucial therapeutic strategy. Interestingly, synthetic STAT3 inhibitors could not manage to enter clinical trials due to their short half-lives, poor bioavailability and non-specific effects. Here, we have shown that Galiellalactone, a natural, irreversible STAT3 inhibitor, blocks IL-6 and WNT5A-mediated STAT3 Phosphorylation and migration in BRAF-inhibitor (BRAFi)-sensitive melanoma cells. Galiellalactone treatment leads to prolonged STAT3 inhibition, which obstructs both metastatic functions and secretome-mediated paracrine interactions in both BRAFi-sensitive and resistant melanoma cells, via the reduction of MCAM and N-cadherin protein expression. Altogether, our findings support that Galiellalactone is a potential inhibitor that could block the metastatic function of drug-resistant melanoma cells and also obstruct the possible paracrine activation of STAT3 conferred by stromal cells present in the tumor microenvironment.

STAT3在转移性和治疗抵抗性黑色素瘤中的过度激活通过促进肿瘤-基质相互作用驱动肿瘤发生和免疫治疗抵抗，使其抑制成为关键的治疗策略。有趣的是，合成STAT3抑制剂由于半衰期短、生物利用度差和非特异性作用而无法进入临床试验。在这里，我们已经证明Galiellalactone，一种天然的，不可逆的STAT3抑制剂，阻断IL-6和wnt5a介导的STAT3在braf抑制剂（BRAFi）敏感黑色素瘤细胞中的磷酸化和迁移。Galiellalactone治疗导致STAT3抑制延长，通过降低MCAM和N-cadherin蛋白表达，阻断brfi敏感和耐药黑色素瘤细胞的转移功能和分泌组介导的旁分泌相互作用。总之，我们的研究结果支持加利利内酯是一种潜在的抑制剂，可以阻断耐药黑色素瘤细胞的转移功能，也可以阻断肿瘤微环境中基质细胞可能赋予的STAT3的旁分泌激活。

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引用次数: 0

Histone methyltransferase NSD3 orchestrates early erythropoiesis by regulating erythroid progenitor cell differentiation and survival 组蛋白甲基转移酶NSD3通过调节红细胞祖细胞的分化和存活来调控早期红细胞生成。

IF 3.7 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2025-12-06 DOI: 10.1016/j.bbamcr.2025.120095

Arunim Shah , Shobhita Katiyar , Chandra Prakash Chaturvedi , Bhuvnesh Rai , Khaliqur Rahman , Naresh Tripathy , Sanjeev Yadav , Anshul Gupta

Histone methyltransferase NSD3 also known KMT3F, is an epigenetic regulator that methylates histone H3 at lysine 36, a mark associated with gene activation. Functionally, NSD3 is crucial for supporting various cellular and developmental processes, and its deregulation can lead to various disease conditions, including solid and hematological cancers. However, the role of NSD3 in regulating normal hematopoiesis remains elusive. In the present study, we have investigated the role of NSD3 in regulating human erythropoiesis, the process of red blood cell production from hematopoietic stem cells (HSCs). To achieve this, we isolated CD34⁺ HSCs from a healthy donor and subjected them to ex-vivo erythroid differentiation. Using a combination of lentiviral transduction and small hairpin RNA-mediated knockdown approaches, we targeted NSD3 depletion during the early phases of erythroid differentiation. Our results demonstrated that depleting NSD3 on days 2 and 7 of ex-vivo erythroid differentiation resulted in notable disruptions of erythropoiesis process. Specifically, NSD3 downregulation resulted in a decreased megakaryocyte-erythroid progenitors, reduced colony formation, and a significant decrease in erythroid differentiation markers. Furthermore, NSD3 depletion altered erythroid differentiation by favouring basophilic erythroblasts over ortho/polychromatic erythrocytes. At the transcriptomic level, NSD3 depletion led to the downregulation of key hematopoiesis-specific transcription factors and genes associated with erythroid differentiation and hemoglobin synthesis. Additionally, NSD3 depletion also induced apoptosis and hindered cell proliferation, accompanied by altered expression of genes involved in these pathways. Our findings uncover a previously undescribed role of histone methyltransferase NSD3 in regulating human erythropoiesis.

组蛋白甲基转移酶NSD3也被称为KMT3F，是一种表观遗传调节剂，可在赖氨酸36位点甲基化组蛋白H3，赖氨酸36是与基因激活相关的标记。在功能上，NSD3对支持各种细胞和发育过程至关重要，其失调可导致各种疾病，包括实体癌和血液癌。然而，NSD3在调节正常造血中的作用尚不清楚。在本研究中，我们研究了NSD3在调节人红细胞生成中的作用，即造血干细胞（hsc）生成红细胞的过程。为了实现这一目标，我们从健康供体中分离出CD34+造血干细胞，并对其进行离体红细胞分化。结合慢病毒转导和小发夹rna介导的敲低方法，我们针对红细胞分化早期阶段的NSD3缺失。我们的研究结果表明，在离体红细胞分化的第2天和第7天消耗NSD3会导致红细胞生成过程明显中断。具体来说，NSD3下调导致巨核细胞-红系祖细胞减少，集落形成减少，红系分化标志物显著减少。此外，NSD3缺失改变了红细胞分化，使嗜碱性红细胞比正色/多色红细胞更倾向于嗜碱性红细胞。在转录组学水平上，NSD3缺失导致关键造血特异性转录因子和与红细胞分化和血红蛋白合成相关的基因下调。此外，NSD3缺失还会诱导细胞凋亡和阻碍细胞增殖，并伴有这些途径相关基因的表达改变。我们的发现揭示了先前未描述的组蛋白甲基转移酶NSD3在调节人类红细胞生成中的作用。

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引用次数: 0

Activation of YBX1 and JAK2/STAT3 pathways by RIOK1 increases lenvatinib resistance in hepatocellular carcinoma cells RIOK1激活YBX1和JAK2/STAT3通路可增加肝癌细胞对lenvatinib的耐药性。

IF 3.7 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2025-12-05 DOI: 10.1016/j.bbamcr.2025.120094

Ruirui Cao , Yue Wu , Yawei Li , Mengting Shi , Tiantian Zhang , Weijie Sun , Yang Liu , Rui Wang

Hepatocellular carcinoma (HCC) is fatal and does not respond well to classical chemotherapies. This paper investigates the molecular mechanism of RIOK1 on lenvatinib resistance in HCC cells. Differentially expressed genes were identified before and after lenvatinib treatment using transcriptome sequencing. Lenvatinib resistance in HCC cells was detected. The interaction between RIOK1 and YBX1 was assessed. YBX1, JAK2, and STAT3 phosphorylation levels were detected. The effect of YBX1 and JAK2/STAT3 pathway activator on lenvatinib resistance was analyzed. Tumor volume and weight, and apoptosis were analyzed in lenvatinib-treated nude mice. RIOK1, YBX1, and JAK2/STAT3 were reduced in HCC cells after lenvatinib treatment. RIOK1 knockdown prevented HCC cell growth and reduced lenvatinib resistance. The interaction between RIOK1 and YBX1 induced Ser 165 phosphorylation, thereby promoting nuclear localization of YBX1. YBX1, JAK2, and STAT3 phosphorylation levels were elevated upon RIOK1 overexpression. YBX1 overexpression and JAK2/STAT3 pathway activator mitigated the anticancer effect of RIOK1 knockdown and increased lenvatinib resistance. Tumor volume, apoptosis, KI67, YBX1, and JAK2/STAT3 phosphorylation levels were reduced in tumor tissue after RIOK1 knockdown and increased after further YBX1 overexpression. Overall, RIOK1 activates the JAK2/STAT3 pathway by promoting YBX1 phosphorylation, leading to HCC progression and lenvatinib resistance.

肝细胞癌（HCC）是致命的，对传统的化疗没有很好的反应。本文探讨RIOK1在肝癌细胞lenvatinib耐药中的分子机制。利用转录组测序方法鉴定lenvatinib治疗前后的差异表达基因。检测肝癌细胞对Lenvatinib的耐药性。评估RIOK1与YBX1的相互作用。检测YBX1、JAK2和STAT3磷酸化水平。分析YBX1和JAK2/STAT3通路激活因子对lenvatinib耐药的影响。对lenvatinib处理裸鼠的肿瘤体积、重量及细胞凋亡进行分析。lenvatinib治疗后，肝癌细胞中的RIOK1、YBX1和JAK2/STAT3减少。RIOK1敲低可阻止HCC细胞生长并降低lenvatinib耐药性。RIOK1与YBX1的相互作用诱导Ser 165磷酸化，从而促进YBX1的核定位。YBX1、JAK2和STAT3的磷酸化水平在RIOK1过表达时升高。YBX1过表达和JAK2/STAT3通路激活因子减轻了RIOK1敲低的抗癌作用，增加了lenvatinib耐药性。RIOK1敲低后，肿瘤组织中肿瘤体积、凋亡、KI67、YBX1和JAK2/STAT3磷酸化水平降低，YBX1过表达后，肿瘤组织中YBX1磷酸化水平升高。总的来说，RIOK1通过促进YBX1磷酸化激活JAK2/STAT3通路，导致HCC进展和lenvatinib耐药。

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引用次数: 0