Pub Date : 2024-06-18DOI: 10.1126/scisignal.adr0297
Wei Wong
Suppression of GLP-1 release by a gut microbiota–derived metabolite induces polycystic ovary syndrome.
肠道微生物群衍生代谢物抑制 GLP-1 的释放会诱发多囊卵巢综合征。
{"title":"A gut punch for PCOS","authors":"Wei Wong","doi":"10.1126/scisignal.adr0297","DOIUrl":"10.1126/scisignal.adr0297","url":null,"abstract":"<div >Suppression of GLP-1 release by a gut microbiota–derived metabolite induces polycystic ovary syndrome.</div>","PeriodicalId":21658,"journal":{"name":"Science Signaling","volume":null,"pages":null},"PeriodicalIF":7.3,"publicationDate":"2024-06-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141421618","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-06-18DOI: 10.1126/scisignal.adi4747
Shane C. Wright, Charlotte Avet, Supriya A. Gaitonde, Itziar Muneta-Arrate, Christian Le Gouill, Mireille Hogue, Billy Breton, Stefania Koutsilieri, Rebeca Diez Alarcia, Madeleine Héroux, Volker M. Lauschke, Michel Bouvier
G protein–coupled receptors (GPCRs) regulate cellular signaling processes by coupling to diverse combinations of heterotrimeric G proteins composed of Gα, Gβ, and Gγ subunits. Biosensors based on bioluminescence resonance energy transfer (BRET) have advanced our understanding of GPCR functional selectivity. Some BRET biosensors monitor ligand-induced conformational changes in the receptor or G proteins, whereas others monitor the recruitment of downstream effectors to sites of G protein activation. Here, we compared the ability of conformation-and activation-based BRET biosensors to assess the coupling of various class A and B GPCRs to specific Gα proteins in cultured cells. These GPCRs included serotonin 5-HT2A and 5-HT7 receptors, the GLP-1 receptor (GLP-1R), and the M3 muscarinic receptor. We observed different signaling profiles between the two types of sensors, highlighting how data interpretation could be affected by the nature of the biosensor. We also found that the identity of the Gβγ subunits used in the assay could differentially influence the selectivity of a receptor toward Gα subtypes, emphasizing the importance of the receptor-Gβγ pairing in determining Gα coupling specificity. Last, the addition of epitope tags to the receptor could affect stoichiometry and coupling selectivity and yield artifactual findings. These results highlight the need for careful sensor selection and experimental design when probing GPCR–G protein coupling.
G 蛋白偶联受体（GPCR）通过与由 Gα、Gβ 和 Gγ 亚基组成的异三聚体 G 蛋白的不同组合耦合来调节细胞信号传导过程。基于生物发光共振能量转移（BRET）的生物传感器推进了我们对 GPCR 功能选择性的了解。一些 BRET 生物传感器监测配体诱导的受体或 G 蛋白构象变化，而另一些则监测下游效应物招募到 G 蛋白活化位点的情况。在这里，我们比较了基于构象和活化的 BRET 生物传感器评估培养细胞中各种 A 类和 B 类 GPCR 与特定 Gα 蛋白耦合的能力。这些 GPCR 包括血清素 5-HT2A 和 5-HT7 受体、GLP-1 受体（GLP-1R）和 M3 肌卡因受体。我们观察到两类传感器的信号特征各不相同，这突出说明了数据解读可能会受到生物传感器性质的影响。我们还发现，检测中使用的 Gβγ 亚基的特性会不同程度地影响受体对 Gα 亚型的选择性，这强调了受体-Gβγ 配对在决定 Gα 偶联特异性方面的重要性。最后，在受体上添加表位标签可能会影响化学计量和耦合选择性，并产生伪造的结果。这些结果突出表明，在探测 GPCR-G 蛋白耦合时，需要仔细选择传感器和设计实验。
{"title":"Conformation- and activation-based BRET sensors differentially report on GPCR–G protein coupling","authors":"Shane C. Wright, Charlotte Avet, Supriya A. Gaitonde, Itziar Muneta-Arrate, Christian Le Gouill, Mireille Hogue, Billy Breton, Stefania Koutsilieri, Rebeca Diez Alarcia, Madeleine Héroux, Volker M. Lauschke, Michel Bouvier","doi":"10.1126/scisignal.adi4747","DOIUrl":"10.1126/scisignal.adi4747","url":null,"abstract":"<div >G protein–coupled receptors (GPCRs) regulate cellular signaling processes by coupling to diverse combinations of heterotrimeric G proteins composed of Gα, Gβ, and Gγ subunits. Biosensors based on bioluminescence resonance energy transfer (BRET) have advanced our understanding of GPCR functional selectivity. Some BRET biosensors monitor ligand-induced conformational changes in the receptor or G proteins, whereas others monitor the recruitment of downstream effectors to sites of G protein activation. Here, we compared the ability of conformation-and activation-based BRET biosensors to assess the coupling of various class A and B GPCRs to specific Gα proteins in cultured cells. These GPCRs included serotonin 5-HT<sub>2A</sub> and 5-HT<sub>7</sub> receptors, the GLP-1 receptor (GLP-1R), and the M<sub>3</sub> muscarinic receptor. We observed different signaling profiles between the two types of sensors, highlighting how data interpretation could be affected by the nature of the biosensor. We also found that the identity of the Gβγ subunits used in the assay could differentially influence the selectivity of a receptor toward Gα subtypes, emphasizing the importance of the receptor-Gβγ pairing in determining Gα coupling specificity. Last, the addition of epitope tags to the receptor could affect stoichiometry and coupling selectivity and yield artifactual findings. These results highlight the need for careful sensor selection and experimental design when probing GPCR–G protein coupling.</div>","PeriodicalId":21658,"journal":{"name":"Science Signaling","volume":null,"pages":null},"PeriodicalIF":7.3,"publicationDate":"2024-06-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.science.org/doi/reader/10.1126/scisignal.adi4747","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141421619","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-06-18DOI: 10.1126/scisignal.adi8743
Kun Ping Lu, Xiao Zhen Zhou
Protein phosphorylation is one of the most common mechanisms regulating cellular signaling pathways, and many kinases and phosphatases are proven drug targets. Upon phosphorylation, protein functions can be further regulated by the distinct isomerase Pin1 through cis-trans isomerization. Numerous protein targets and many important roles have now been elucidated for Pin1. However, no tools are available to detect or target cis and trans conformation events in cells. The development of Pin1 inhibitors and stereo- and phospho-specific antibodies has revealed that cis and trans conformations have distinct and often opposing cellular functions. Aberrant conformational changes due to the dysregulation of Pin1 can drive pathogenesis but can be effectively targeted in age-related diseases, including cancers and neurodegenerative disorders. Here, we review advances in understanding the roles of Pin1 signaling in health and disease and highlight conformational regulation as a distinct signal transduction checkpoint in disease development and treatment.
{"title":"Pin1-catalyzed conformational regulation after phosphorylation: A distinct checkpoint in cell signaling and drug discovery","authors":"Kun Ping Lu, Xiao Zhen Zhou","doi":"10.1126/scisignal.adi8743","DOIUrl":"10.1126/scisignal.adi8743","url":null,"abstract":"<div >Protein phosphorylation is one of the most common mechanisms regulating cellular signaling pathways, and many kinases and phosphatases are proven drug targets. Upon phosphorylation, protein functions can be further regulated by the distinct isomerase Pin1 through cis-trans isomerization. Numerous protein targets and many important roles have now been elucidated for Pin1. However, no tools are available to detect or target cis and trans conformation events in cells. The development of Pin1 inhibitors and stereo- and phospho-specific antibodies has revealed that cis and trans conformations have distinct and often opposing cellular functions. Aberrant conformational changes due to the dysregulation of Pin1 can drive pathogenesis but can be effectively targeted in age-related diseases, including cancers and neurodegenerative disorders. Here, we review advances in understanding the roles of Pin1 signaling in health and disease and highlight conformational regulation as a distinct signal transduction checkpoint in disease development and treatment.</div>","PeriodicalId":21658,"journal":{"name":"Science Signaling","volume":null,"pages":null},"PeriodicalIF":7.3,"publicationDate":"2024-06-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141421620","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-06-11DOI: 10.1126/scisignal.adn8376
Vivian Chua, Melisa Lopez-Anton, Mizue Terai, Ryota Tanaka, Usman Baqai, Timothy J. Purwin, Jelan I. Haj, Francis J. Waltrich Jr., Isabella Trachtenberg, Kristine Luo, Rohith Tudi, Angela Jeon, Anna Han, Inna Chervoneva, Michael A. Davies, Julio A. Aguirre-Ghiso, Takami Sato, Andrew E. Aplin
Uveal melanoma (UM) is the deadliest form of eye cancer in adults. Inactivating mutations and/or loss of expression of the gene encoding BRCA1-associated protein 1 (BAP1) in UM tumors are associated with an increased risk of metastasis. To investigate the mechanisms underlying this risk, we explored the functional consequences of BAP1 deficiency. UM cell lines expressing mutant BAP1 grew more slowly than those expressing wild-type BAP1 in culture and in vivo. The ability of BAP1 reconstitution to restore cell proliferation in BAP1-deficient cells required its deubiquitylase activity. Proteomic analysis showed that BAP1-deficient cells had decreased phosphorylation of ribosomal S6 and its upstream regulator, p70S6K1, compared with both wild-type and BAP1 reconstituted cells. In turn, expression of p70S6K1 increased S6 phosphorylation and proliferation of BAP1-deficient UM cells. Consistent with these findings, BAP1 mutant primary UM tumors expressed lower amounts of p70S6K1 target genes, and S6 phosphorylation was decreased in BAP1 mutant patient-derived xenografts (PDXs), which grew more slowly than wild-type PDXs in the liver (the main metastatic site of UM) in mice. BAP1-deficient UM cells were also more resistant to amino acid starvation, which was associated with diminished phosphorylation of S6. These studies demonstrate that BAP1 deficiency slows the proliferation of UM cells through regulation of S6 phosphorylation. These characteristics may be associated with metastasis by ensuring survival during amino acid starvation.
葡萄膜黑色素瘤（UM）是成人中最致命的眼癌。UM肿瘤中编码BRCA1相关蛋白1（BAP1）基因的失活突变和/或表达缺失与转移风险增加有关。为了研究这种风险的内在机制，我们探讨了 BAP1 缺乏的功能性后果。在培养和体内，表达突变型 BAP1 的 UM 细胞系比表达野生型 BAP1 的细胞系生长更慢。在 BAP1 缺乏的细胞中，BAP1 重组恢复细胞增殖的能力需要其去泛素化酶活性。蛋白质组分析表明，与野生型细胞和BAP1重组细胞相比，BAP1缺陷细胞中核糖体S6及其上游调节因子p70S6K1的磷酸化减少。反过来，p70S6K1 的表达增加了 S6 磷酸化和 BAP1 缺陷 UM 细胞的增殖。与这些发现相一致的是，BAP1突变型原发性UM肿瘤表达的p70S6K1靶基因数量较低，BAP1突变型患者衍生异种移植物（PDXs）中的S6磷酸化也降低了，在小鼠肝脏（UM的主要转移部位）中，PDXs的生长速度比野生型PDXs慢。缺乏 BAP1 的 UM 细胞对氨基酸饥饿的抵抗力也更强，这与 S6 磷酸化减少有关。这些研究表明，BAP1 缺乏会通过调节 S6 磷酸化减缓 UM 细胞的增殖。这些特征可能与转移有关，因为它们能确保细胞在氨基酸饥饿期间存活。
{"title":"Slow proliferation of BAP1-deficient uveal melanoma cells is associated with reduced S6 signaling and resistance to nutrient stress","authors":"Vivian Chua, Melisa Lopez-Anton, Mizue Terai, Ryota Tanaka, Usman Baqai, Timothy J. Purwin, Jelan I. Haj, Francis J. Waltrich Jr., Isabella Trachtenberg, Kristine Luo, Rohith Tudi, Angela Jeon, Anna Han, Inna Chervoneva, Michael A. Davies, Julio A. Aguirre-Ghiso, Takami Sato, Andrew E. Aplin","doi":"10.1126/scisignal.adn8376","DOIUrl":"10.1126/scisignal.adn8376","url":null,"abstract":"<div >Uveal melanoma (UM) is the deadliest form of eye cancer in adults. Inactivating mutations and/or loss of expression of the gene encoding BRCA1-associated protein 1 (BAP1) in UM tumors are associated with an increased risk of metastasis. To investigate the mechanisms underlying this risk, we explored the functional consequences of BAP1 deficiency. UM cell lines expressing mutant <i>BAP1</i> grew more slowly than those expressing wild-type <i>BAP1</i> in culture and in vivo. The ability of BAP1 reconstitution to restore cell proliferation in BAP1-deficient cells required its deubiquitylase activity. Proteomic analysis showed that BAP1-deficient cells had decreased phosphorylation of ribosomal S6 and its upstream regulator, p70S6K1, compared with both wild-type and BAP1 reconstituted cells. In turn, expression of p70S6K1 increased S6 phosphorylation and proliferation of BAP1-deficient UM cells. Consistent with these findings, <i>BAP1</i> mutant primary UM tumors expressed lower amounts of p70S6K1 target genes, and S6 phosphorylation was decreased in <i>BAP1</i> mutant patient-derived xenografts (PDXs), which grew more slowly than wild-type PDXs in the liver (the main metastatic site of UM) in mice. BAP1-deficient UM cells were also more resistant to amino acid starvation, which was associated with diminished phosphorylation of S6. These studies demonstrate that BAP1 deficiency slows the proliferation of UM cells through regulation of S6 phosphorylation. These characteristics may be associated with metastasis by ensuring survival during amino acid starvation.</div>","PeriodicalId":21658,"journal":{"name":"Science Signaling","volume":null,"pages":null},"PeriodicalIF":7.3,"publicationDate":"2024-06-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141307192","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Neuroendocrine prostate cancer (PCa) (NEPC), an aggressive subtype that is associated with poor prognosis, may arise after androgen deprivation therapy (ADT). We investigated the molecular mechanisms by which ADT induces neuroendocrine differentiation in advanced PCa. We found that transmembrane protein 1 (MCTP1), which has putative Ca2+ sensing function and multiple Ca2+-binding C2 domains, was abundant in samples from patients with advanced PCa. MCTP1 was associated with the expression of the EMT-associated transcription factors ZBTB46, FOXA2, and HIF1A. The increased abundance of MCTP1 promoted PC3 prostate cancer cell migration and neuroendocrine differentiation and was associated with SNAI1-dependent EMT in C4-2 PCa cells after ADT. ZBTB46 interacted with FOXA2 and HIF1A and increased the abundance of MCTP1 in a hypoxia-dependent manner. MCTP1 stimulated Ca2+ signaling and AKT activation to promote EMT and neuroendocrine differentiation by increasing the SNAI1-dependent expression of EMT and neuroendocrine markers, effects that were blocked by knockdown of MCTP1. These data suggest an oncogenic role for MCTP1 in the maintenance of a rare and aggressive prostate cancer subtype through its response to Ca2+ and suggest its potential as a therapeutic target.
{"title":"MCTP1 increases the malignancy of androgen-deprived prostate cancer cells by inducing neuroendocrine differentiation and EMT","authors":"Yen-Nien Liu, Wei-Yu Chen, Hsiu-Lien Yeh, Wei-Hao Chen, Kuo-Ching Jiang, Han-Ru Li, Phan Vu Thuy Dung, Zi-Qing Chen, Wei-Jiunn Lee, Michael Hsiao, Jiaoti Huang, Yu-Ching Wen","doi":"10.1126/scisignal.adc9142","DOIUrl":"10.1126/scisignal.adc9142","url":null,"abstract":"<div >Neuroendocrine prostate cancer (PCa) (NEPC), an aggressive subtype that is associated with poor prognosis, may arise after androgen deprivation therapy (ADT). We investigated the molecular mechanisms by which ADT induces neuroendocrine differentiation in advanced PCa. We found that transmembrane protein 1 (MCTP1), which has putative Ca<sup>2+</sup> sensing function and multiple Ca<sup>2+</sup>-binding C2 domains, was abundant in samples from patients with advanced PCa. MCTP1 was associated with the expression of the EMT-associated transcription factors ZBTB46, FOXA2, and HIF1A. The increased abundance of MCTP1 promoted PC3 prostate cancer cell migration and neuroendocrine differentiation and was associated with SNAI1-dependent EMT in C4-2 PCa cells after ADT. ZBTB46 interacted with FOXA2 and HIF1A and increased the abundance of MCTP1 in a hypoxia-dependent manner. MCTP1 stimulated Ca<sup>2+</sup> signaling and AKT activation to promote EMT and neuroendocrine differentiation by increasing the SNAI1-dependent expression of EMT and neuroendocrine markers, effects that were blocked by knockdown of MCTP1. These data suggest an oncogenic role for MCTP1 in the maintenance of a rare and aggressive prostate cancer subtype through its response to Ca<sup>2+</sup> and suggest its potential as a therapeutic target.</div>","PeriodicalId":21658,"journal":{"name":"Science Signaling","volume":null,"pages":null},"PeriodicalIF":7.3,"publicationDate":"2024-06-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141307191","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-06-11DOI: 10.1126/scisignal.adq9088
Amy E. Baek
Apoptosis of immature peripheral B cells may be due to a lack of survival signals rather than clonal deletion.
未成熟外周 B 细胞的凋亡可能是由于缺乏生存信号，而不是克隆性缺失。
{"title":"To B or not to B","authors":"Amy E. Baek","doi":"10.1126/scisignal.adq9088","DOIUrl":"10.1126/scisignal.adq9088","url":null,"abstract":"<div >Apoptosis of immature peripheral B cells may be due to a lack of survival signals rather than clonal deletion.</div>","PeriodicalId":21658,"journal":{"name":"Science Signaling","volume":null,"pages":null},"PeriodicalIF":7.3,"publicationDate":"2024-06-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141307193","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
A long-standing question in the field of signal transduction is how distinct signaling pathways interact with each other to control cell behavior. Growth factor receptors and G protein–coupled receptors (GPCRs) are the two major signaling hubs in eukaryotes. Given that the mechanisms by which they signal independently have been extensively characterized, we investigated how they may cross-talk with each other. Using linear ion trap mass spectrometry and cell-based biophysical, biochemical, and phenotypic assays, we found at least three distinct ways in which epidermal growth factor affected canonical G protein signaling by the Gi-coupled GPCR CXCR4 through the phosphorylation of Gαi. Phosphomimicking mutations in two residues in the αE helix of Gαi (tyrosine-154/tyrosine-155) suppressed agonist-induced Gαi activation while promoting constitutive Gβγ signaling. Phosphomimicking mutations in the P loop (serine-44, serine-47, and threonine-48) suppressed Gi activation entirely, thus completely segregating growth factor and GPCR pathways. As expected, most of the phosphorylation events appeared to affect intrinsic properties of Gαi proteins, including conformational stability, nucleotide binding, and the ability to associate with and to release Gβγ. However, one phosphomimicking mutation, targeting the carboxyl-terminal residue tyrosine-320, promoted mislocalization of Gαi from the plasma membrane, a previously uncharacterized mechanism of suppressing GPCR signaling through G protein subcellular compartmentalization. Together, these findings elucidate not only how growth factor and chemokine signals cross-talk through the phosphorylation-dependent modulation of Gαi but also how such cross-talk may generate signal diversity.
信号转导领域一个长期存在的问题是，不同的信号通路是如何相互作用来控制细胞行为的。生长因子受体和 G 蛋白偶联受体（GPCR）是真核生物的两大信号枢纽。鉴于它们独立发出信号的机制已被广泛表征，我们研究了它们如何相互交叉。利用线性离子阱质谱法和基于细胞的生物物理、生物化学和表型测定，我们发现表皮生长因子通过 Gαi 磷酸化影响 Gi 偶联 GPCR CXCR4 的典型 G 蛋白信号传导至少有三种不同的方式。Gαi的αE螺旋中两个残基（酪氨酸-154/酪氨酸-155）的磷酸化突变抑制了激动剂诱导的Gαi激活，同时促进了组成型Gβγ信号传导。P 环（丝氨酸-44、丝氨酸-47 和苏氨酸-48）的磷酸化突变完全抑制了 Gi 的激活，从而完全分离了生长因子和 GPCR 途径。不出所料，大多数磷酸化事件似乎都会影响 Gαi 蛋白的固有特性，包括构象稳定性、核苷酸结合以及与 Gβγ 结合和释放 Gβγ 的能力。然而，一个以羧基末端残基酪氨酸-320 为靶点的磷酸模仿突变促进了 Gαi 从质膜上的错误定位，这是一种以前未曾描述过的通过 G 蛋白亚细胞区隔抑制 GPCR 信号转导的机制。总之，这些发现不仅阐明了生长因子和趋化因子信号如何通过磷酸化依赖性调节 Gαi 相互传递，而且还阐明了这种相互传递如何产生信号多样性。
{"title":"Growth factor–dependent phosphorylation of Gαi shapes canonical signaling by G protein–coupled receptors","authors":"Suchismita Roy, Saptarshi Sinha, Ananta James Silas, Majid Ghassemian, Irina Kufareva, Pradipta Ghosh","doi":"10.1126/scisignal.ade8041","DOIUrl":"10.1126/scisignal.ade8041","url":null,"abstract":"<div >A long-standing question in the field of signal transduction is how distinct signaling pathways interact with each other to control cell behavior. Growth factor receptors and G protein–coupled receptors (GPCRs) are the two major signaling hubs in eukaryotes. Given that the mechanisms by which they signal independently have been extensively characterized, we investigated how they may cross-talk with each other. Using linear ion trap mass spectrometry and cell-based biophysical, biochemical, and phenotypic assays, we found at least three distinct ways in which epidermal growth factor affected canonical G protein signaling by the G<sub>i</sub>-coupled GPCR CXCR4 through the phosphorylation of Gα<sub>i</sub>. Phosphomimicking mutations in two residues in the α<sub>E</sub> helix of Gα<sub>i</sub> (tyrosine-154/tyrosine-155) suppressed agonist-induced Gα<sub>i</sub> activation while promoting constitutive Gβγ signaling. Phosphomimicking mutations in the P loop (serine-44, serine-47, and threonine-48) suppressed G<sub>i</sub> activation entirely, thus completely segregating growth factor and GPCR pathways. As expected, most of the phosphorylation events appeared to affect intrinsic properties of Gα<sub>i</sub> proteins, including conformational stability, nucleotide binding, and the ability to associate with and to release Gβγ. However, one phosphomimicking mutation, targeting the carboxyl-terminal residue tyrosine-320, promoted mislocalization of Gα<sub>i</sub> from the plasma membrane, a previously uncharacterized mechanism of suppressing GPCR signaling through G protein subcellular compartmentalization. Together, these findings elucidate not only how growth factor and chemokine signals cross-talk through the phosphorylation-dependent modulation of Gα<sub>i</sub> but also how such cross-talk may generate signal diversity.</div>","PeriodicalId":21658,"journal":{"name":"Science Signaling","volume":null,"pages":null},"PeriodicalIF":7.3,"publicationDate":"2024-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141249013","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-05-28DOI: 10.1126/scisignal.ado6266
Lavinia Palamiuc, Jared L. Johnson, Zeinab Haratipour, Ryan M. Loughran, Woong Jae Choi, Gurpreet K. Arora, Vivian Tieu, Kyanh Ly, Alicia Llorente, Sophia Crabtree, Jenny C. Y. Wong, Archna Ravi, Thorsten Wiederhold, Rabi Murad, Raymond D. Blind, Brooke M. Emerling
Phosphoinositides are essential signaling molecules. The PI5P4K family of phosphoinositide kinases and their substrates and products, PI5P and PI4,5P2, respectively, are emerging as intracellular metabolic and stress sensors. We performed an unbiased screen to investigate the signals that these kinases relay and the specific upstream regulators controlling this signaling node. We found that the core Hippo pathway kinases MST1/2 phosphorylated PI5P4Ks and inhibited their signaling in vitro and in cells. We further showed that PI5P4K activity regulated several Hippo- and YAP-related phenotypes, specifically decreasing the interaction between the key Hippo proteins MOB1 and LATS and stimulating the YAP-mediated genetic program governing epithelial-to-mesenchymal transition. Mechanistically, we showed that PI5P interacted with MOB1 and enhanced its interaction with LATS, thereby providing a signaling connection between the Hippo pathway and PI5P4Ks. These findings reveal how these two important evolutionarily conserved signaling pathways are integrated to regulate metazoan development and human disease.
{"title":"Hippo and PI5P4K signaling intersect to control the transcriptional activation of YAP","authors":"Lavinia Palamiuc, Jared L. Johnson, Zeinab Haratipour, Ryan M. Loughran, Woong Jae Choi, Gurpreet K. Arora, Vivian Tieu, Kyanh Ly, Alicia Llorente, Sophia Crabtree, Jenny C. Y. Wong, Archna Ravi, Thorsten Wiederhold, Rabi Murad, Raymond D. Blind, Brooke M. Emerling","doi":"10.1126/scisignal.ado6266","DOIUrl":"10.1126/scisignal.ado6266","url":null,"abstract":"<div >Phosphoinositides are essential signaling molecules. The PI5P4K family of phosphoinositide kinases and their substrates and products, PI5P and PI4,5P<sub>2</sub>, respectively, are emerging as intracellular metabolic and stress sensors. We performed an unbiased screen to investigate the signals that these kinases relay and the specific upstream regulators controlling this signaling node. We found that the core Hippo pathway kinases MST1/2 phosphorylated PI5P4Ks and inhibited their signaling in vitro and in cells. We further showed that PI5P4K activity regulated several Hippo- and YAP-related phenotypes, specifically decreasing the interaction between the key Hippo proteins MOB1 and LATS and stimulating the YAP-mediated genetic program governing epithelial-to-mesenchymal transition. Mechanistically, we showed that PI5P interacted with MOB1 and enhanced its interaction with LATS, thereby providing a signaling connection between the Hippo pathway and PI5P4Ks. These findings reveal how these two important evolutionarily conserved signaling pathways are integrated to regulate metazoan development and human disease.</div>","PeriodicalId":21658,"journal":{"name":"Science Signaling","volume":null,"pages":null},"PeriodicalIF":7.3,"publicationDate":"2024-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141162494","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-05-28DOI: 10.1126/scisignal.adq5728
Leslie K. Ferrarelli
Astrocyte-secreted neurocan guides the formation of inhibitory circuits in the brain.
星形胶质细胞分泌的神经蛋白能引导大脑抑制回路的形成。
{"title":"Glia in inhibitory synaptogenesis","authors":"Leslie K. Ferrarelli","doi":"10.1126/scisignal.adq5728","DOIUrl":"10.1126/scisignal.adq5728","url":null,"abstract":"<div >Astrocyte-secreted neurocan guides the formation of inhibitory circuits in the brain.</div>","PeriodicalId":21658,"journal":{"name":"Science Signaling","volume":null,"pages":null},"PeriodicalIF":7.3,"publicationDate":"2024-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141162488","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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