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Chromatin protein complexes involved in gene repression in lamina-associated domains. 参与板层相关结构域基因抑制的染色质蛋白复合物
IF 9.4 1区 生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-11-01 Epub Date: 2024-09-25 DOI: 10.1038/s44318-024-00214-1
Stefano G Manzo, Abdelghani Mazouzi, Christ Leemans, Tom van Schaik, Nadia Neyazi, Marjon S van Ruiten, Benjamin D Rowland, Thijn R Brummelkamp, Bas van Steensel

Lamina-associated domains (LADs) are large chromatin regions that are associated with the nuclear lamina (NL) and form a repressive environment for transcription. The molecular players that mediate gene repression in LADs are currently unknown. Here, we performed FACS-based whole-genome genetic screens in human cells using LAD-integrated fluorescent reporters to identify such regulators. Surprisingly, the screen identified very few NL proteins, but revealed roles for dozens of known chromatin regulators. Among these are the negative elongation factor (NELF) complex and interacting factors involved in RNA polymerase pausing, suggesting that regulation of transcription elongation is a mechanism to repress transcription in LADs. Furthermore, the chromatin remodeler complex BAF and the activation complex Mediator can work both as activators and repressors in LADs, depending on the local context and possibly by rewiring heterochromatin. Our data indicate that the fundamental regulators of transcription and chromatin remodeling, rather than interaction with NL proteins, play a major role in transcription regulation within LADs.

核薄层相关域(LAD)是与核薄层(NL)相关的大型染色质区域,形成了抑制转录的环境。目前,LADs 中介导基因抑制的分子角色尚不清楚。在这里,我们使用 LAD 整合荧光报告物对人类细胞进行了基于 FACS 的全基因组遗传筛选,以确定此类调控因子。令人惊讶的是,筛选结果发现的 NL 蛋白很少,但却揭示了数十种已知染色质调控因子的作用。其中包括负伸长因子(NELF)复合物和参与 RNA 聚合酶暂停的相互作用因子,这表明转录伸长的调控是 LADs 中抑制转录的一种机制。此外,染色质重塑复合物 BAF 和激活复合物 Mediator 在 LADs 中既可作为激活因子,也可作为抑制因子,这取决于局部环境,也可能是通过重新连接异染色质来实现的。我们的数据表明,转录和染色质重塑的基本调节因子,而不是与 NL 蛋白的相互作用,在 LADs 内的转录调节中发挥着重要作用。
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引用次数: 0
RNA fine-tunes estrogen receptor-alpha binding on low-affinity DNA motifs for transcriptional regulation. RNA 可微调雌激素受体-α 与低亲和性 DNA 基团的结合,从而实现转录调控。
IF 9.4 1区 生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-11-01 Epub Date: 2024-09-16 DOI: 10.1038/s44318-024-00225-y
Deepanshu Soota, Bharath Saravanan, Rajat Mann, Tripti Kharbanda, Dimple Notani

Transcription factors (TFs) regulate gene expression by binding with varying strengths to DNA via their DNA-binding domain. Additionally, some TFs also interact with RNA, which modulates transcription factor binding to chromatin. However, whether RNA-mediated TF binding results in differential transcriptional outcomes remains unknown. In this study, we demonstrate that estrogen receptor α (ERα), a ligand-activated TF, interacts with RNA in a ligand-dependent manner. Defects in RNA binding lead to genome-wide loss of ERα recruitment, particularly at weaker ERα-motifs. Furthermore, ERα mobility in the nucleus increases in the absence of its RNA-binding capacity. Unexpectedly, this increased mobility coincides with robust polymerase loading and transcription of ERα-regulated genes that harbor low-strength motifs. However, highly stable binding of ERα on chromatin negatively impacts ligand-dependent transcription. Collectively, our results suggest that RNA interactions spatially confine ERα on low-affinity sites to fine-tune gene transcription.

转录因子(TF)通过其 DNA 结合域以不同强度与 DNA 结合,从而调节基因表达。此外,一些转录因子还与 RNA 相互作用,从而调节转录因子与染色质的结合。然而,RNA 介导的 TF 结合是否会导致不同的转录结果仍是未知数。在这项研究中,我们证明了雌激素受体α(ERα)是一种配体激活的TF,它以配体依赖的方式与RNA相互作用。RNA 结合缺陷会导致全基因组范围的 ERα 招募丧失,尤其是在较弱的 ERα 位点。此外,在缺乏 RNA 结合能力的情况下,ERα 在细胞核中的流动性也会增加。意想不到的是,ERα移动性增加的同时,ERα调控基因的聚合酶加载和转录能力也很强,而ERα调控基因中含有低强度的基序。然而,ERα在染色质上的高度稳定结合会对配体依赖性转录产生负面影响。总之,我们的研究结果表明,RNA相互作用在空间上将ERα限制在低亲和力位点上,以微调基因转录。
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引用次数: 0
Combining systems and synthetic biology for in vivo enzymology. 将系统生物学与合成生物学相结合,用于体内酶学研究。
IF 9.4 1区 生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-11-01 Epub Date: 2024-09-25 DOI: 10.1038/s44318-024-00251-w
Sara Castaño-Cerezo, Alexandre Chamas, Hanna Kulyk, Christian Treitz, Floriant Bellvert, Andreas Tholey, Virginie Galéote, Carole Camarasa, Stéphanie Heux, Luis F Garcia-Alles, Pierre Millard, Gilles Truan

Enzymatic parameters are classically determined in vitro, under conditions that are far from those encountered in cells, casting doubt on their physiological relevance. We developed a generic approach combining tools from synthetic and systems biology to measure enzymatic parameters in vivo. In the context of a synthetic carotenoid pathway in Saccharomyces cerevisiae, we focused on a phytoene synthase and three phytoene desaturases, which are difficult to study in vitro. We designed, built, and analyzed a collection of yeast strains mimicking substantial variations in substrate concentration by strategically manipulating the expression of geranyl-geranyl pyrophosphate (GGPP) synthase. We successfully determined in vivo Michaelis-Menten parameters (KM, Vmax, and kcat) for GGPP-converting phytoene synthase from absolute metabolomics, fluxomics and proteomics data, highlighting differences between in vivo and in vitro parameters. Leveraging the versatility of the same set of strains, we then extracted enzymatic parameters for two of the three phytoene desaturases. Our approach demonstrates the feasibility of assessing enzymatic parameters directly in vivo, providing a novel perspective on the kinetic characteristics of enzymes in real cellular conditions.

酶学参数通常是在体外测定的,其测定条件与细胞内的条件相差甚远,这使人们对其生理相关性产生怀疑。我们开发了一种结合合成生物学和系统生物学工具的通用方法,用于测量体内的酶参数。在类胡萝卜素合成途径的背景下,我们重点研究了一种植物烯合成酶和三种植物烯去饱和酶,这些酶在体外很难研究。我们设计、构建并分析了一系列酵母菌株,通过策略性地操纵牻牛儿基-牻牛儿基焦磷酸(GGPP)合成酶的表达,模拟底物浓度的巨大变化。我们成功地从绝对代谢组学、通量组学和蛋白质组学数据中确定了GGPP转化植物烯合成酶的体内Michaelis-Menten参数(KM、Vmax和kcat),突出了体内和体外参数之间的差异。利用同一组菌株的多功能性,我们随后提取了三种植物烯去饱和酶中两种酶的酶学参数。我们的方法证明了直接评估体内酶参数的可行性,为了解真实细胞条件下酶的动力学特性提供了一个新的视角。
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引用次数: 0
Real-time assessment of mitochondrial DNA heteroplasmy dynamics at the single-cell level. 在单细胞水平上实时评估线粒体 DNA 异构动态。
IF 9.4 1区 生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-11-01 Epub Date: 2024-08-05 DOI: 10.1038/s44318-024-00183-5
Rodaria Roussou, Dirk Metzler, Francesco Padovani, Felix Thoma, Rebecca Schwarz, Boris Shraiman, Kurt M Schmoller, Christof Osman

Mitochondrial DNA (mtDNA) is present in multiple copies within cells and is required for mitochondrial ATP generation. Even within individual cells, mtDNA copies can differ in their sequence, a state known as heteroplasmy. The principles underlying dynamic changes in the degree of heteroplasmy remain incompletely understood, due to the inability to monitor this phenomenon in real time. Here, we employ mtDNA-based fluorescent markers, microfluidics, and automated cell tracking, to follow mtDNA variants in live heteroplasmic yeast populations at the single-cell level. This approach, in combination with direct mtDNA tracking and data-driven mathematical modeling reveals asymmetric partitioning of mtDNA copies during cell division, as well as limited mitochondrial fusion and fission frequencies, as critical driving forces for mtDNA variant segregation. Given that our approach also facilitates assessment of segregation between intact and mutant mtDNA, we anticipate that it will be instrumental in elucidating the mechanisms underlying the purifying selection of mtDNA.

线粒体 DNA(mtDNA)在细胞内有多个拷贝,是线粒体 ATP 生成所必需的。即使在单个细胞内,mtDNA拷贝的序列也可能不同,这种状态被称为异质性。由于无法对这一现象进行实时监测,人们对异质性程度动态变化的基本原理仍然不甚了解。在这里,我们采用基于 mtDNA 的荧光标记、微流控技术和自动细胞追踪技术,在单细胞水平上追踪活的异质酵母群体中的 mtDNA 变异。这种方法与直接 mtDNA 跟踪和数据驱动的数学建模相结合,揭示了细胞分裂过程中 mtDNA 拷贝的非对称分割,以及线粒体融合和分裂频率的限制,是 mtDNA 变异分离的关键驱动力。鉴于我们的方法还有助于评估完整 mtDNA 和突变 mtDNA 之间的分离,我们预计它将有助于阐明 mtDNA 纯化选择的内在机制。
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引用次数: 0
Non-cell-autonomous regulation of germline proteostasis by insulin/IGF-1 signaling-induced dietary peptide uptake via PEPT-1. 胰岛素/IGF-1 信号通过 PEPT-1 诱导的膳食肽吸收对生殖细胞蛋白稳态的非细胞自主调节
IF 9.4 1区 生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-11-01 Epub Date: 2024-09-16 DOI: 10.1038/s44318-024-00234-x
Tahir Muhammad, Stacey L Edwards, Allison C Morphis, Mary V Johnson, Vitor De Oliveira, Tomasz Chamera, Siyan Liu, Ngoc Gia Tuong Nguyen, Jian Li

Gametogenesis involves active protein synthesis and is proposed to rely on proteostasis. Our previous work in C. elegans indicates that germline development requires coordinated activities of insulin/IGF-1 signaling (IIS) and HSF-1, the central regulator of the heat shock response. However, the downstream mechanisms were not identified. Here, we show that depletion of HSF-1 from germ cells impairs chaperone gene expression, causing protein degradation and aggregation and, consequently, reduced fecundity and gamete quality. Conversely, reduced IIS confers germ cell resilience to HSF-1 depletion-induced protein folding defects and various proteotoxic stresses. Surprisingly, this effect was not mediated by an enhanced stress response, which underlies longevity in low IIS conditions, but by reduced ribosome biogenesis and translation rate. We found that IIS activates the expression of intestinal peptide transporter PEPT-1 by alleviating its repression by FOXO/DAF-16, allowing dietary proteins to be efficiently incorporated into an amino acid pool that fuels germline protein synthesis. Our data suggest this non-cell-autonomous pathway is critical for proteostasis regulation during gametogenesis.

生殖细胞的形成涉及活跃的蛋白质合成,并被认为依赖于蛋白稳态。我们之前在 elegans 中的研究表明,生殖细胞的发育需要胰岛素/IGF-1 信号传导(IIS)和热休克反应的核心调节因子 HSF-1 的协调活动。然而,其下游机制尚未确定。在这里,我们发现生殖细胞中 HSF-1 的耗竭会损害伴侣蛋白基因的表达,导致蛋白质降解和聚集,从而降低生殖力和配子质量。相反,IIS的减少使生殖细胞对HSF-1耗竭引起的蛋白质折叠缺陷和各种蛋白毒性应激具有恢复能力。令人惊讶的是,这种效应并不是通过应激反应的增强(这是低 IIS 条件下长寿的基础)介导的,而是通过核糖体生物生成和翻译速率的降低介导的。我们发现,IIS通过减轻肠肽转运体PEPT-1受FOXO/DAF-16的抑制,激活了它的表达,使膳食蛋白质有效地融入氨基酸池,从而促进种系蛋白质的合成。我们的数据表明,这种非细胞自主途径对配子发生过程中的蛋白稳态调节至关重要。
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引用次数: 0
Cancer cell-intrinsic biosynthesis of itaconate promotes tumor immunogenicity. 癌细胞内在的伊他康酸生物合成促进了肿瘤免疫原性。
IF 9.4 1区 生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-11-01 Epub Date: 2024-09-30 DOI: 10.1038/s44318-024-00217-y
Zining Wang, Lei Cui, Yanxun Lin, Bitao Huo, Hongxia Zhang, Chunyuan Xie, Huanling Zhang, Yongxiang Liu, Huan Jin, Hui Guo, Mengyun Li, Xiaojuan Wang, Penghui Zhou, Peng Huang, Jinyun Liu, Xiaojun Xia

The Krebs cycle byproduct itaconate has recently emerged as an important metabolite regulating macrophage immune functions, but its role in tumor cells remains unknown. Here, we show that increased tumor-intrinsic cis-aconitate decarboxylase (ACOD1 or CAD, encoded by immune-responsive gene 1, Irg1) expression and itaconate production promote tumor immunogenicity and anti-tumor immune responses. Furthermore, we identify thimerosal, a vaccine preservative, as a specific inducer of IRG1 expression in tumor cells but not in macrophages, thereby enhancing tumor immunogenicity. Mechanistically, thimerosal induces itaconate production through a ROS-RIPK3-IRF1 signaling axis in tumor cells. Further, increased IRG1/itaconate upregulates antigen presentation-related gene expression via promoting TFEB nuclear translocation. Intratumoral injection of thimerosal induced itaconate production, activated the tumor immune microenvironment, and inhibited tumor growth in a T cell-dependent manner. Importantly, IRG1 deficiency markedly impaired tumor response to thimerosal treatment. Furthermore, itaconate induction by thimerosal potentiates the anti-tumor efficacy of adoptive T-cell therapy and anti-PD1 therapy in a mouse lymphoma model. Hence, our findings identify a new role for tumor intrinsic IRG1/itaconate in promoting tumor immunogenicity and provide a translational means to increase immunotherapy efficacy.

最近,克雷布斯循环副产物伊他康酸已成为调节巨噬细胞免疫功能的重要代谢物,但它在肿瘤细胞中的作用仍然未知。在这里,我们发现肿瘤内在顺式-乌头酸脱羧酶(ACOD1 或 CAD,由免疫反应基因 1 Irg1 编码)表达的增加和伊塔康酸的产生促进了肿瘤的免疫原性和抗肿瘤免疫反应。此外,我们还发现疫苗防腐剂硫柳汞是肿瘤细胞中 IRG1 表达的特异性诱导剂,而不是巨噬细胞中的特异性诱导剂,从而增强了肿瘤的免疫原性。从机理上讲,硫柳汞通过肿瘤细胞中的 ROS-RIPK3-IRF1 信号轴诱导伊它康酸的产生。此外,IRG1/伊他康酸的增加会通过促进 TFEB 核易位来上调抗原递呈相关基因的表达。瘤内注射硫柳汞可诱导伊他康酸的产生,激活肿瘤免疫微环境,并以T细胞依赖的方式抑制肿瘤生长。重要的是,IRG1的缺乏会明显降低肿瘤对硫柳汞治疗的反应。此外,在小鼠淋巴瘤模型中,硫柳汞诱导的伊他康酸能增强采用T细胞疗法和抗PD1疗法的抗肿瘤疗效。因此,我们的研究结果确定了肿瘤内在 IRG1/itaconate 在促进肿瘤免疫原性方面的新作用,并为提高免疫疗法疗效提供了一种转化手段。
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引用次数: 0
A human progeria-associated BAF-1 mutation modulates gene expression and accelerates aging in C. elegans. 人类早衰症相关的 BAF-1 基因突变会调节基因表达并加速优雅小鼠的衰老。
IF 9.4 1区 生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-11-01 Epub Date: 2024-10-04 DOI: 10.1038/s44318-024-00261-8
Raquel Romero-Bueno, Adrián Fragoso-Luna, Cristina Ayuso, Nina Mellmann, Alan Kavsek, Christian G Riedel, Jordan D Ward, Peter Askjaer

Alterations in the nuclear envelope are linked to a variety of rare diseases termed laminopathies. A single amino acid substitution at position 12 (A12T) of the human nuclear envelope protein BAF (Barrier to Autointegration Factor) causes Néstor-Guillermo Progeria Syndrome (NGPS). This premature ageing condition leads to growth retardation and severe skeletal defects, but the underlying mechanisms are unknown. Here, we have generated a novel in vivo model for NGPS by modifying the baf-1 locus in C. elegans to mimic the human NGPS mutation. These baf-1(G12T) mutant worms displayed multiple phenotypes related to fertility, lifespan, and stress resistance. Importantly, nuclear morphology deteriorated faster during aging in baf-1(G12T) compared to wild-type animals, recapitulating an important hallmark of cells from progeria patients. Although localization of BAF-1(G12T) was similar to wild-type BAF-1, lamin accumulation at the nuclear envelope was reduced in mutant worms. Tissue-specific chromatin binding and transcriptome analyses showed reduced BAF-1 association in most genes deregulated by the baf-1(G12T) mutation, suggesting that altered BAF chromatin association induces NGPS phenotypes via altered gene expression.

核包膜的改变与多种罕见疾病有关,这些疾病被称为片层病。人类核包膜蛋白 BAF(自结合屏障因子)第 12 位的一个氨基酸置换(A12T)导致了内斯托-吉列尔莫早衰综合症(NGPS)。这种早衰症会导致生长迟缓和严重的骨骼缺陷,但其潜在机制尚不清楚。在这里,我们通过修改优雅子的 baf-1 基因座来模拟人类 NGPS 基因突变,从而产生了一种新型的 NGPS 体内模型。这些 baf-1(G12T)突变体蠕虫表现出与生育能力、寿命和抗应激能力有关的多种表型。重要的是,与野生型动物相比,baf-1(G12T)的核形态在衰老过程中退化得更快,这再现了早衰症患者细胞的一个重要特征。虽然BAF-1(G12T)的定位与野生型BAF-1相似,但在突变体蠕虫中,核膜上的片层积累减少了。组织特异性染色质结合和转录组分析表明,baf-1(G12T)突变导致大多数基因的BAF-1关联性降低,这表明BAF染色质关联性的改变是通过改变基因表达来诱导NGPS表型的。
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引用次数: 0
IF1 is a cold-regulated switch of ATP synthase hydrolytic activity to support thermogenesis in brown fat. IF1 是 ATP 合成酶水解活性的冷调节开关,支持棕色脂肪的热生成。
IF 9.4 1区 生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-11-01 Epub Date: 2024-09-16 DOI: 10.1038/s44318-024-00215-0
Henver S Brunetta, Anna S Jung, Fernando Valdivieso-Rivera, Stepheny C de Campos Zani, Joel Guerra, Vanessa O Furino, Annelise Francisco, Marcelo Berçot, Pedro M Moraes-Vieira, Susanne Keipert, Martin Jastroch, Laurent O Martinez, Carlos H Sponton, Roger F Castilho, Marcelo A Mori, Alexander Bartelt

While mechanisms controlling uncoupling protein-1 (UCP1) in thermogenic adipocytes play a pivotal role in non-shivering thermogenesis, it remains unclear whether F1Fo-ATP synthase function is also regulated in brown adipose tissue (BAT). Here, we show that inhibitory factor 1 (IF1, encoded by Atp5if1), an inhibitor of ATP synthase hydrolytic activity, is a critical negative regulator of brown adipocyte energy metabolism. In vivo, IF1 levels are diminished in BAT of cold-adapted mice compared to controls. Additionally, the capacity of ATP synthase to generate mitochondrial membrane potential (MMP) through ATP hydrolysis (the so-called "reverse mode" of ATP synthase) is increased in brown fat. In cultured brown adipocytes, IF1 overexpression results in an inability of mitochondria to sustain the MMP upon adrenergic stimulation, leading to a quiescent-like phenotype in brown adipocytes. In mice, adeno-associated virus-mediated IF1 overexpression in BAT suppresses adrenergic-stimulated thermogenesis and decreases mitochondrial respiration in BAT. Taken together, our work identifies downregulation of IF1 upon cold as a critical event for the facilitation of the reverse mode of ATP synthase as well as to enable energetic adaptation of BAT to effectively support non-shivering thermogenesis.

虽然控制产热脂肪细胞中解偶联蛋白-1(UCP1)的机制在非颤抖性产热中起着关键作用,但棕色脂肪组织(BAT)中的 F1Fo-ATP 合酶功能是否也受到调控仍不清楚。在这里,我们发现抑制因子 1(IF1,由 Atp5if1 编码)是 ATP 合酶水解活性的抑制剂,是棕色脂肪细胞能量代谢的关键负调控因子。与对照组相比,冷适应小鼠体内 BAT 中的 IF1 水平降低。此外,棕色脂肪中的 ATP 合成酶通过 ATP 水解产生线粒体膜电位(MMP)的能力(即所谓的 ATP 合成酶 "反向模式")也有所增加。在培养的棕色脂肪细胞中,IF1 过表达会导致线粒体在肾上腺素能刺激下无法维持 MMP,从而导致棕色脂肪细胞出现类似静止的表型。在小鼠体内,腺相关病毒介导的 IF1 在 BAT 中的过表达抑制了肾上腺素能刺激下的产热,并降低了 BAT 的线粒体呼吸。综上所述,我们的研究发现,IF1 在寒冷时的下调是促进 ATP 合成酶反向模式的关键事件,也是使 BAT 进行能量适应以有效支持非颤抖性产热的关键事件。
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引用次数: 0
Ezrin, radixin, and moesin are dispensable for macrophage migration and cellular cortex mechanics. Ezrin、radixin 和 moesin 对于巨噬细胞迁移和细胞皮层力学是不可或缺的。
IF 9.4 1区 生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-11-01 Epub Date: 2024-07-18 DOI: 10.1038/s44318-024-00173-7
Perrine Verdys, Javier Rey Barroso, Adeline Girel, Joseph Vermeil, Martin Bergert, Thibaut Sanchez, Arnaud Métais, Thomas Mangeat, Elisabeth Bellard, Claire Bigot, Catherine Astarie-Dequeker, Arnaud Labrousse, Jean-Philippe Girard, Isabelle Maridonneau-Parini, Christel Vérollet, Frédéric Lagarrigue, Alba Diz-Muñoz, Julien Heuvingh, Matthieu Piel, Olivia du Roure, Véronique Le Cabec, Sébastien Carréno, Renaud Poincloux

The cellular cortex provides crucial mechanical support and plays critical roles during cell division and migration. The proteins of the ERM family, comprised of ezrin, radixin, and moesin, are central to these processes by linking the plasma membrane to the actin cytoskeleton. To investigate the contributions of the ERM proteins to leukocyte migration, we generated single and triple ERM knockout macrophages. Surprisingly, we found that even in the absence of ERM proteins, macrophages still form the different actin structures promoting cell migration, such as filopodia, lamellipodia, podosomes, and ruffles. Furthermore, we discovered that, unlike every other cell type previously investigated, the single or triple knockout of ERM proteins does not affect macrophage migration in diverse contexts. Finally, we demonstrated that the loss of ERMs in macrophages does not affect the mechanical properties of their cortex. These findings challenge the notion that ERMs are universally essential for cortex mechanics and cell migration and support the notion that the macrophage cortex may have diverged from that of other cells to allow for their uniquely adaptive cortical plasticity.

细胞皮层提供重要的机械支持,并在细胞分裂和迁移过程中发挥关键作用。由 ezrin、radixin 和 moesin 组成的 ERM 家族蛋白通过连接质膜和肌动蛋白细胞骨架,在这些过程中发挥着核心作用。为了研究ERM蛋白对白细胞迁移的贡献,我们产生了单个和三个ERM基因敲除的巨噬细胞。令人惊讶的是,我们发现即使在缺乏ERM蛋白的情况下,巨噬细胞仍能形成促进细胞迁移的不同肌动蛋白结构,如丝状体、片状体、荚膜和褶皱。此外,我们还发现,与之前研究过的其他细胞类型不同,单次或三次敲除ERM蛋白不会影响巨噬细胞在不同环境下的迁移。最后,我们证明巨噬细胞中 ERM 的缺失不会影响其皮质的机械特性。这些发现质疑了ERMs对皮质力学和细胞迁移普遍至关重要的观点,并支持了巨噬细胞皮质可能与其他细胞的皮质不同,从而使其具有独特的适应性皮质可塑性的观点。
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引用次数: 0
Carboxy-terminal polyglutamylation regulates signaling and phase separation of the Dishevelled protein. 羧基末端多谷氨酰化调节 Dishevelled 蛋白的信号传递和相分离。
IF 9.4 1区 生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-11-01 Epub Date: 2024-09-30 DOI: 10.1038/s44318-024-00254-7
Marek Kravec, Ondrej Šedo, Jana Nedvědová, Miroslav Micka, Marie Šulcová, Nikodém Zezula, Kristína Gömöryová, David Potěšil, Ranjani Sri Ganji, Sara Bologna, Igor Červenka, Zbyněk Zdráhal, Jakub Harnoš, Konstantinos Tripsianes, Carsten Janke, Cyril Bařinka, Vítězslav Bryja

Polyglutamylation is a reversible posttranslational modification that is catalyzed by enzymes of the tubulin tyrosine ligase-like (TTLL) family. Here, we found that TTLL11 generates a previously unknown type of polyglutamylation that is initiated by the addition of a glutamate residue to the free C-terminal carboxyl group of a substrate protein. TTLL11 efficiently polyglutamylates the Wnt signaling protein Dishevelled 3 (DVL3), thereby changing the interactome of DVL3. Polyglutamylation increases the capacity of DVL3 to get phosphorylated, to undergo phase separation, and to act in the noncanonical Wnt pathway. Both carboxy-terminal polyglutamylation and the resulting reduction in phase separation capacity of DVL3 can be reverted by the deglutamylating enzyme CCP6, demonstrating a causal relationship between TTLL11-mediated polyglutamylation and phase separation. Thus, C-terminal polyglutamylation represents a new type of posttranslational modification, broadening the range of proteins that can be modified by polyglutamylation and providing the first evidence that polyglutamylation can modulate protein phase separation.

多谷氨酰化是一种可逆的翻译后修饰,由类管突酪氨酸连接酶(TTLL)家族的酶催化。在这里,我们发现 TTLL11 会产生一种以前未知的多谷氨酰化,这种多谷氨酰化是通过将谷氨酸残基添加到底物蛋白质游离的 C 端羧基上而启动的。TTLL11 能有效地多谷氨酰化 Wnt 信号蛋白 Dishevelled 3 (DVL3),从而改变 DVL3 的相互作用组。多聚戊酰胺化提高了 DVL3 被磷酸化、发生相分离以及在非经典 Wnt 通路中发挥作用的能力。羧基末端多聚戊酰胺化和由此导致的 DVL3 相分离能力下降都能被脱戊酰胺酶 CCP6 所逆转,这表明 TTLL11 介导的多聚戊酰胺化与相分离之间存在因果关系。因此,C 端聚谷氨酰化是一种新型的翻译后修饰,它拓宽了可被聚谷氨酰化修饰的蛋白质范围,并首次证明了聚谷氨酰化可调节蛋白质的相分离。
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