Science Signaling最新文献

A sequencing-based screening method identifies regulators of EGFR signaling from nonviable mutants in Caenorhabditis elegans

IF 6.7 1区生物学

Science Signaling Pub Date : 2025-02-25

Hillel T. Schwartz, Paul W. Sternberg

{"title":"A sequencing-based screening method identifies regulators of EGFR signaling from nonviable mutants in Caenorhabditis elegans","authors":"Hillel T. Schwartz, Paul W. Sternberg","doi":"","DOIUrl":"","url":null,"abstract":"<div >Suppressor screens can identify genetic modifiers of biochemical pathways but generally require that the suppressed mutant be viable and fertile. We developed a screening method that obviated this requirement and enabled the identification of mutations that partially suppressed the early developmental arrest and lethality caused by loss of the epidermal growth factor (EGF) receptor ortholog LET-23 in Caenorhabditis elegans. We chemically mutagenized animals carrying the loss-of-function allele let-23(sy15), recovered let-23(sy15) homozygotes that escaped early developmental arrest but were nevertheless inviable, and sequenced their genomes. Testing of candidate causal mutations identified 11 genes that, when mutated, mitigated the early lethality caused by loss of EGF signaling. These included genes encoding homologs of the small guanosine triphosphatase (GTPase) Ras (let-60), which is a downstream effector of LET-23, and of regulators of the small GTPase Rho, including the homolog of the phosphotyrosine-binding protein TENSIN (tns-1). We also recovered suppressing mutations in genes encoding nuclear proteins that protect against DNA damage, including the homolog of MutS homolog 4 (him-14). Genetic experiments were consistent with the repression of Rho activity or the activation of the DNA damage response compensating for the loss of EGF signaling. This sequencing-based, whole-animal screening method may be adapted to other organisms to enable the identification of mutations for which the phenotype does not allow the recovery of viable animals.</div>","PeriodicalId":21658,"journal":{"name":"Science Signaling","volume":"18 875","pages":""},"PeriodicalIF":6.7,"publicationDate":"2025-02-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143490070","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}

引用次数: 0

Glutamatergic argonaute2 promotes the formation of the neurovascular unit in mice

IF 6.7 1区生物学

Science Signaling Pub Date : 2025-02-25

Chandan Sona, Yu-Te Yeh, Yunxiao Li, Xiaoxuan Liu, Adhideb Ghosh, Laura C. Hinte, Min-Chi Ku, Thomas Rathjen, Thoralf Niendorf, Guoxing Yu, Shiqi Jia, Natalia L. Kononenko, Andreas Hermann, Jiankai Luo, Juntang Lin, Ferdinand von Meyenn, Xin Yan, Matthew N. Poy

{"title":"Glutamatergic argonaute2 promotes the formation of the neurovascular unit in mice","authors":"Chandan Sona, Yu-Te Yeh, Yunxiao Li, Xiaoxuan Liu, Adhideb Ghosh, Laura C. Hinte, Min-Chi Ku, Thomas Rathjen, Thoralf Niendorf, Guoxing Yu, Shiqi Jia, Natalia L. Kononenko, Andreas Hermann, Jiankai Luo, Juntang Lin, Ferdinand von Meyenn, Xin Yan, Matthew N. Poy","doi":"","DOIUrl":"","url":null,"abstract":"<div >Proper formation of the complex neurovascular unit (NVU) along with the blood-brain barrier is critical for building and sustaining a healthy, functioning central nervous system. The RNA binding protein argonaute2 (Ago2) mediates microRNA (miRNA)–mediated gene silencing, which is critical for many facets of brain development, including NVU development. Here, we found that Ago2 in glutamatergic neurons was critical for NVU formation in the developing cortices of mice. Glutamatergic neuron–specific loss of Ago2 diminished synaptic formation, neuronal-to-endothelial cell contacts, and morphogenesis of the brain vasculature, ultimately compromising the integrity of the blood-brain barrier. Ago2 facilitated miRNA targeting of phosphatase and tensin homolog (Pten) mRNA, which encodes a phosphatase that modulates reelin-dependent phosphatidylinositol 3-kinase (PI3K)–Akt signaling within the glutamatergic subpopulation. Conditionally deleting Pten in Ago2-deficient neurons restored Akt2 phosphorylation as well as postnatal development and survival. Several mutations in AGO2 impair small RNA silencing and are associated with Lessel-Kreienkamp syndrome, a neurodevelopmental disorder. When expressed in a neuronal cell line, these human AGO2 loss-of-function variants failed to suppress PTEN, resulting in attenuated PI3K-Akt signaling, further indicating that dysregulation of Ago2 function may contribute to both impaired development and neurological disorders. Together, these results identify Ago2 as central to the engagement of neurons with blood vessels in the developing brain.</div>","PeriodicalId":21658,"journal":{"name":"Science Signaling","volume":"18 875","pages":""},"PeriodicalIF":6.7,"publicationDate":"2025-02-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143490069","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}

引用次数: 0

Dulling the sweet tooth

IF 6.7 1区生物学

Science Signaling Pub Date : 2025-02-25

Annalisa M. VanHook

引用次数: 0

Stem cell health from mom’s gut

IF 6.7 1区生物学

Science Signaling Pub Date : 2025-02-18

Leslie K. Ferrarelli

引用次数: 0

Sustained chromosomal passenger complex activity preserves the pluripotency of human embryonic carcinoma cells

IF 6.7 1区生物学

Science Signaling Pub Date : 2025-02-18

Takaaki Tsunematsu, Yasuhiro Mouri, Wenhua Shao, Rieko Arakaki, Jan G. Ruppert, Kensaku Murano, Naozumi Ishimaru, Daniele Guardavaccaro, Michele Pagano, Yasusei Kudo

{"title":"Sustained chromosomal passenger complex activity preserves the pluripotency of human embryonic carcinoma cells","authors":"Takaaki Tsunematsu, Yasuhiro Mouri, Wenhua Shao, Rieko Arakaki, Jan G. Ruppert, Kensaku Murano, Naozumi Ishimaru, Daniele Guardavaccaro, Michele Pagano, Yasusei Kudo","doi":"","DOIUrl":"","url":null,"abstract":"<div >Human embryonic carcinoma (hEC) cells are derived from teratocarcinomas, exhibit robust proliferation, have a high differentiation potential, are the malignant counterparts of human embryonic stem cells (hESCs), and are considered hESC-like. The chromosomal passenger complex (CPC), made up of the microtuble binding protein Borealin, the kinase Aurora-B, the CPC-stabilizing inner centromere protein (INCENP), and the inhibitor of apoptosis family member Survivin, regulates cell division and is active exclusively during mitosis in somatic cells. The anaphase-promoting complex/cyclosome and its cofactor Cdh1 (APC/CCdh1) is a ubiquitylating complex that catalyzes the degradation of Aurora-B and Borealin in somatic cells but has low activity during interphase in hESCs. Here, we found that Borealin and Aurora-B exhibited sustained stability throughout the cell cycle of hEC cells due to low APC/CCdh1 activity. In contrast with somatic cells, CPC activity persisted across the cell cycle of hEC cells because of diminished APC/CCdh1 activity. Disrupting the CPC complex by depleting its constituents triggered spontaneous differentiation in hEC cells. As hEC cells differentiated, APC/CCdh1 activation curtailed CPC activity. Inactivating the CPC by pharmacologically inhibiting Aurora-B induced hEC cell differentiation by activating the epithelial-to-mesenchymal transition (EMT) program. Hence, APC/CCdh1-mediated termination of CPC activity triggered hEC cell differentiation. Collectively, these findings demonstrate a role for the CPC in governing hESC cell fate.</div>","PeriodicalId":21658,"journal":{"name":"Science Signaling","volume":"18 874","pages":""},"PeriodicalIF":6.7,"publicationDate":"2025-02-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143439263","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}

引用次数: 0

Sustained Gαs signaling mediated by vasopressin type 2 receptors is ligand dependent but endocytosis and β-arrestin independent

IF 6.7 1区生物学

Science Signaling Pub Date : 2025-02-18

Larissa B. Teixeira, Marie-José Blouin, Christian Le Gouill, Louis-Philippe Picard, Claudio M. Costa-Neto, Michel Bouvier, Lucas T. Parreiras-e-Silva

{"title":"Sustained Gαs signaling mediated by vasopressin type 2 receptors is ligand dependent but endocytosis and β-arrestin independent","authors":"Larissa B. Teixeira, Marie-José Blouin, Christian Le Gouill, Louis-Philippe Picard, Claudio M. Costa-Neto, Michel Bouvier, Lucas T. Parreiras-e-Silva","doi":"","DOIUrl":"","url":null,"abstract":"<div >The canonical model of G protein–coupled receptor (GPCR) signaling comprises G protein activation at the plasma membrane, followed by receptor phosphorylation and β-arrestin recruitment, which leads to receptor desensitization and endocytosis. However, the activation of some GPCRs results in sustained G protein signaling from intracellular compartments in a manner reportedly dependent on β-arrestin and receptor endocytosis. The vasopressin type 2 receptor (V2R) can be activated by two structurally similar hormones, arginine vasopressin and oxytocin, both of which stimulate the production of the second messenger cyclic adenosine monophosphate (cAMP). In this study, we showed that sustained V2R signaling and endosomal Gαs (stimulatory G protein alpha subunit) translocation could occur without β-arrestin–mediated receptor endocytosis and was primarily controlled by the residence time of the ligand on the receptor. β-Arrestin had opposing effects on sustained signaling: It facilitated receptor internalization into endosomes, where it activated Gαs, and promoted cAMP production from this compartment. However, β-arrestin–mediated receptor endocytosis also induced ligand dissociation due to the acidic endosomal environment, thereby limiting the signal. Overall, our data suggest that signals originating at the plasma membrane play a dominant role in sustained V2R signaling stimulated by arginine vasopressin.</div>","PeriodicalId":21658,"journal":{"name":"Science Signaling","volume":"18 874","pages":""},"PeriodicalIF":6.7,"publicationDate":"2025-02-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.science.org/doi/reader/10.1126/scisignal.adf6206","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143439262","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}

引用次数: 0

Seeing new partners for GPCRs

IF 6.7 1区生物学

Science Signaling Pub Date : 2025-02-11

John F. Foley

引用次数: 0

Targeting c-MYC and gain-of-function p53 through inhibition or degradation of the kinase LZK suppresses the growth of HNSCC tumors

IF 6.7 1区生物学

Science Signaling Pub Date : 2025-02-11

Amy L. Funk, Meghri Katerji, Marwa Afifi, Katherine Nyswaner, Carolyn C. Woodroofe, Zoe C. Edwards, Eric Lindberg, Knickole L. Bergman, Nancy R. Gough, Maxine R. Rubin, Kamila Karpińska, Eleanor W. Trotter, Sweta Dash, Amy L. Ries, Amy James, Christina M. Robinson, Simone Difilippantonio, Baktiar O. Karim, Ting-Chia Chang, Li Chen, Xin Xu, James H. Doroshow, Ivan Ahel, Anna A. Marusiak, Rolf E. Swenson, Steven D. Cappell, John Brognard

{"title":"Targeting c-MYC and gain-of-function p53 through inhibition or degradation of the kinase LZK suppresses the growth of HNSCC tumors","authors":"Amy L. Funk, Meghri Katerji, Marwa Afifi, Katherine Nyswaner, Carolyn C. Woodroofe, Zoe C. Edwards, Eric Lindberg, Knickole L. Bergman, Nancy R. Gough, Maxine R. Rubin, Kamila Karpińska, Eleanor W. Trotter, Sweta Dash, Amy L. Ries, Amy James, Christina M. Robinson, Simone Difilippantonio, Baktiar O. Karim, Ting-Chia Chang, Li Chen, Xin Xu, James H. Doroshow, Ivan Ahel, Anna A. Marusiak, Rolf E. Swenson, Steven D. Cappell, John Brognard","doi":"","DOIUrl":"","url":null,"abstract":"<div >The worldwide annual frequency and lethality of head and neck squamous cell carcinoma (HNSCC) is not improving, and thus, new therapeutic approaches are needed. Approximately 70% of HNSCC cases have either amplification or overexpression of MAP3K13, which encodes the kinase LZK. Here, we found that LZK is a therapeutic target in HNSCC and that small-molecule inhibition of its catalytic function decreased the viability of HNSCC cells with amplified MAP3K13. Inhibition of LZK suppressed tumor growth in MAP3K13-amplified xenografts derived from HNSCC patients. LZK stabilized the transcription factor c-MYC through its kinase activity and gain-of-function mutants of p53 in a kinase-independent manner. We designed a proteolysis-targeting chimera (PROTAC) that induced LZK degradation, leading to decreased abundance of both c-MYC and gain-of-function p53, and reduced the viability of HNSCC cells. Our findings demonstrate that LZK-targeted therapeutics, particularly PROTACs, may be effective in treating HNSCCs with MAP3K13 amplification.</div>","PeriodicalId":21658,"journal":{"name":"Science Signaling","volume":"18 873","pages":""},"PeriodicalIF":6.7,"publicationDate":"2025-02-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143389482","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}

引用次数: 0

Rebuilding duct work from the gallbladder

IF 6.7 1区生物学

Science Signaling Pub Date : 2025-02-04

Wei Wong

引用次数: 0

G protein peptidomimetics reveal allosteric effects and stepwise interactions in ghrelin receptor–G protein coupling G蛋白多肽模拟物揭示了胃泌素受体-G蛋白耦合中的异构效应和逐步相互作用

IF 6.7 1区生物学

Science Signaling Pub Date : 2025-02-04

Morgane Mannes, Charlotte Martin, Marjorie Damian, Sonia Cantel, Hélène Orcel, Jean-Alain Fehrentz, Bernard Mouillac, Julie Kniazeff, Jean-Louis Banères, Steven Ballet

{"title":"G protein peptidomimetics reveal allosteric effects and stepwise interactions in ghrelin receptor–G protein coupling","authors":"Morgane Mannes, Charlotte Martin, Marjorie Damian, Sonia Cantel, Hélène Orcel, Jean-Alain Fehrentz, Bernard Mouillac, Julie Kniazeff, Jean-Louis Banères, Steven Ballet","doi":"","DOIUrl":"","url":null,"abstract":"<div >G protein–coupled receptor (GPCR) signaling is a dynamic process involving various conformational intermediates in addition to those captured in static three-dimensional structures. Here, we used newly developed G protein peptidomimetics to characterize the interactions of the ghrelin receptor (GHSR) with G proteins. Coupling to the G protein peptidomimetic not only affected the conformational features of the cytoplasmic regions of the receptor where the G protein binds but also allosterically affected the extracellular ligand-binding pocket. These conformational and allosteric changes increased the affinity of G protein–coupled GHSR for the endogenous agonist ghrelin. In addition, our data identified different complexes along the G protein activation pathway that differed in the engagement of the Gαq C-terminal helix. Given that this helix is the main link between the activated receptor and the Gα nucleotide-binding pocket, these findings suggested a stepwise process involving distinct states in GPCR-catalyzed G protein activation. Collectively, our results provide evidence for the dynamic behavior of GPCR–G protein signaling complexes, with such dynamics most likely contributing to signaling selectivity and/or efficacy.</div>","PeriodicalId":21658,"journal":{"name":"Science Signaling","volume":"18 872","pages":""},"PeriodicalIF":6.7,"publicationDate":"2025-02-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143187402","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}

引用次数: 0