18,396 results on '"Active Transport, Cell Nucleus"'
Search Results
2. The interaction between the import carrier Hikeshi and HSP70 is modulated by heat, facilitating the nuclear import of HSP70 under heat stress conditions.
- Author
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Kose S, Yoshioka S, Ogawa Y, Watanabe A, and Imamoto N
- Subjects
- Humans, Cell Nucleus metabolism, Protein Binding, Hot Temperature, HeLa Cells, Nuclear Pore Complex Proteins metabolism, Nuclear Pore Complex Proteins genetics, Mutation, Carrier Proteins, HSP70 Heat-Shock Proteins metabolism, HSP70 Heat-Shock Proteins genetics, Active Transport, Cell Nucleus, Heat-Shock Response
- Abstract
Heat stress strongly triggers the nuclear localization of the molecular chaperone HSP70. Hikeshi functions as a unique nuclear import carrier of HSP70. However, how the nuclear import of HSP70 is activated in response to heat stress remains unclear. Here, we investigated the effects of heat on the nuclear import of HSP70. In vitro transport assays revealed that pretreatment of the test samples with heat facilitated the nuclear import of HSP70. Furthermore, binding of Hikeshi to HSP70 increased when temperatures rose. These results indicated that heat is one of the factors that activates the nuclear import of HSP70. Previous studies showed that the F97A mutation in Hikeshi in an extended loop induced an opening in the hydrophobic pocket and facilitated the translocation of Hikeshi through the nuclear pore complex. We found that nuclear accumulation of HSP70 occurred at a lower temperature in cells expressing the Hikeshi-F97A mutant than in cells expressing wild-type Hikeshi. Collectively, our results show that the movement of the extended loop may play an important role in the interaction of Hikeshi with both FG (phenylalanine-glycine)-nucleoporins and HSP70 in a temperature-dependent manner, resulting in the activation of nuclear import of HSP70 in response to heat stress., (© 2024 Molecular Biology Society of Japan and John Wiley & Sons Australia, Ltd.)
- Published
- 2024
- Full Text
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3. Nuclear export of PML promotes p53-mediated apoptosis and ferroptosis.
- Author
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Ni Y, Chen H, Zhan Q, and Zhuang Z
- Subjects
- Humans, Cell Line, Tumor, Cell Movement, Cell Nucleus metabolism, Membrane Potential, Mitochondrial, Reactive Oxygen Species metabolism, Active Transport, Cell Nucleus, Amino Acid Transport System y+ metabolism, Amino Acid Transport System y+ genetics, Apoptosis, Cell Proliferation, Ferroptosis, Phospholipid Hydroperoxide Glutathione Peroxidase metabolism, Promyelocytic Leukemia Protein metabolism, Promyelocytic Leukemia Protein genetics, Tumor Suppressor Protein p53 metabolism
- Abstract
Promyelocytic leukemia protein (PML), a tumor suppressor protein, plays a key role in cell cycle regulation, apoptosis, senescence and cellular metabolism. Here, we report that PML promotes apoptosis and ferroptosis. Our data showed that PML over-expression inhibited cell proliferation and migration. PML over-expression increased apoptotic cells, nuclear condensation and the loss of mitochondrial membrane potential, accompanied by regulation of Bcl-2 family proteins and reactive oxygen species (ROS) level, suggesting that PML enhanced apoptosis. Meanwhile, PML over-expression not only increased lipid ROS accumulation and Malondialdehyde (MDA) content but also downregulated solute carrier family 7 member 11 (SLC7A11) and glutathione peroxidase 4 (GPX4) expression, indicating that PML enhanced ferroptosis. Additionally, knockdown of p53 attenuated the effect of PML on SLC7A11 and GPX4, and inhibited the increase of lipid ROS and ROS by PML over-expression. Moreover, translocation of PML from nucleus to cytoplasm not only promoted apoptosis and ferroptosis, but also inhibited cell proliferation. Taken together, PML promotes apoptosis and ferroptosis, in which the mediation of p53 and the nuclear export of PML play important roles., Competing Interests: Declaration of competing interest The authors declare no competing interests., (Copyright © 2023. Published by Elsevier Inc.)
- Published
- 2024
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4. FolIws1-driven nuclear translocation of deacetylated FolTFIIS ensures conidiation of Fusarium oxysporum.
- Author
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Qian H, Song L, Wang L, Yang Q, Wu R, Du J, Zheng B, and Liang W
- Subjects
- Acetylation, Plant Diseases microbiology, Cell Nucleus metabolism, Gene Expression Regulation, Fungal, Active Transport, Cell Nucleus, Botrytis genetics, Botrytis metabolism, Botrytis drug effects, Fusarium metabolism, Fusarium drug effects, Fusarium genetics, Fusarium pathogenicity, Spores, Fungal metabolism, Spores, Fungal drug effects, Fungal Proteins metabolism, Fungal Proteins genetics
- Abstract
Plant diseases caused by fungal pathogens pose a great threat to crop production. Conidiation of fungi is critical for disease epidemics and serves as a promising drug target. Here, we show that deacetylation of the FolTFIIS transcription elongation factor is indispensable for Fusarium oxysporum f. sp. lycopersici (Fol) conidiation. Upon microconidiation, Fol decreases K76 acetylation of FolTFIIS by altering the level of controlling enzymes, allowing for its nuclear translocation by FolIws1. Increased nuclear FolTFIIS enhances the transcription of sporulation-related genes and, consequently, enables microconidia production. Deacetylation of FolTFIIS is also critical for the production of macroconidia and chlamydospores, and its homolog has similar functions in Botrytis cinerea. We identify two FolIws1-targeting chemicals that block the conidiation of Fol and have effective activity against a wide range of pathogenic fungi without harm to the hosts. These findings reveal a conserved mechanism of conidiation regulation and provide candidate agrochemicals for disease management., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)
- Published
- 2024
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5. STAT3 Increases CVB3 Replication and Acute Pancreatitis and Myocarditis Pathology via Impeding Nuclear Translocation of STAT1 and Interferon-Stimulated Gene Expression.
- Author
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Liang T, Zhang Z, Bai Z, Xu L, and Xu W
- Subjects
- Animals, Mice, Humans, Coxsackievirus Infections metabolism, Coxsackievirus Infections virology, Coxsackievirus Infections pathology, Coxsackievirus Infections genetics, Cell Nucleus metabolism, Male, Active Transport, Cell Nucleus, Gene Expression Regulation, Acute Disease, Cell Line, Signal Transduction, STAT1 Transcription Factor metabolism, STAT1 Transcription Factor genetics, Myocarditis virology, Myocarditis metabolism, Myocarditis pathology, Myocarditis genetics, STAT3 Transcription Factor metabolism, STAT3 Transcription Factor genetics, Pancreatitis metabolism, Pancreatitis virology, Pancreatitis pathology, Pancreatitis genetics, Enterovirus B, Human physiology, Virus Replication
- Abstract
Acute pancreatitis (AP) is an inflammatory disease initiated by the death of exocrine acinar cells, but its pathogenesis remains unclear. Signal transducer and activator of transcription 3 (STAT3) is a multifunctional factor that regulates immunity and the inflammatory response. The protective role of STAT3 is reported in Coxsackievirus B3 (CVB3)-induced cardiac fibrosis, yet the exact role of STAT3 in modulating viral-induced STAT1 activation and type I interferon (IFN)-stimulated gene (ISG) transcription in the pancreas remains unclarified. In this study, we tested whether STAT3 regulated viral-induced STAT1 translocation. We found that CVB3, particularly capsid VP1 protein, markedly upregulated the phosphorylation and nuclear import of STAT3 (p-STAT3) while it significantly impeded the nuclear translocation of p-STAT1 in the pancreases and hearts of mice on day 3 postinfection (p.i.). Immunoblotting and an immunofluorescent assay demonstrated the increased expression and nuclear translocation of p-STAT3 but a blunted p-STAT1 nuclear translocation in CVB3-infected acinar 266-6 cells. STAT3 shRNA knockdown or STAT3 inhibitors reduced viral replication via the rescue of STAT1 nuclear translocation and increasing the ISRE activity and ISG transcription in vitro. The knockdown of STAT1 blocked the antiviral effect of the STAT3 inhibitor. STAT3 inhibits STAT1 activation by virally inducing a potent inhibitor of IFN signaling, the suppressor of cytokine signaling-3 ((SOCS)-3). Sustained pSTAT1 and the elevated expression of ISGs were induced in SOCS3 knockdown cells. The in vivo administration of HJC0152, a pharmaceutical STAT3 inhibitor, mitigated the viral-induced AP and myocarditis pathology via increasing the IFNβ as well as ISG expression on day 3 p.i. and reducing the viral load in multi-organs. These findings define STAT3 as a negative regulator of the type I IFN response via impeding the nuclear STAT1 translocation that otherwise triggers ISG induction in infected pancreases and hearts. Our findings identify STAT3 as an antagonizing factor of the IFN-STAT1 signaling pathway and provide a potential therapeutic target for viral-induced AP and myocarditis.
- Published
- 2024
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6. Role of charge in enhanced nuclear transport and retention of graphene quantum dots.
- Author
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Gorav G, Khedekar V, Varier GK, and Nandakumar P
- Subjects
- Humans, HeLa Cells, Nuclear Envelope metabolism, Nuclear Pore metabolism, Microscopy, Confocal, Quantum Dots chemistry, Quantum Dots metabolism, Graphite chemistry, Active Transport, Cell Nucleus, Cell Nucleus metabolism
- Abstract
The nuclear pore complexes on the nuclear membrane serve as the exclusive gateway for communication between the nucleus and the cytoplasm, regulating the transport of various molecules, including nucleic acids and proteins. The present work investigates the kinetics of the transport of negatively charged graphene quantum dots through nuclear membranes, focusing on quantifying their transport characteristics. Experiments are carried out in permeabilized HeLa cells using time-lapse confocal fluorescence microscopy. Our findings indicate that negatively charged graphene quantum dots exhibit rapid transport to the nuclei, involving two distinct transport pathways in the translocation process. Complementary experiments on the nuclear import and export of graphene quantum dots validate the bi-directionality of transport, as evidenced by comparable transport rates. The study also shows that the negatively charged graphene quantum dots possess favorable retention properties, underscoring their potential as drug carriers., (© 2024. The Author(s).)
- Published
- 2024
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7. Importin-7-dependent nuclear translocation of the Flavivirus core protein is required for infectious virus production.
- Author
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Itoh Y, Miyamoto Y, Tokunaga M, Suzuki T, Takada A, Ninomiya A, Hishinuma T, Matsuda M, Yoneda Y, Oka M, Suzuki R, Matsuura Y, and Okamoto T
- Subjects
- Humans, Animals, Virus Replication physiology, Viral Core Proteins metabolism, Viral Core Proteins genetics, Karyopherins metabolism, Karyopherins genetics, Flavivirus Infections metabolism, Flavivirus Infections virology, Chlorocebus aethiops, HEK293 Cells, Flavivirus metabolism, Flavivirus physiology, Active Transport, Cell Nucleus, Cell Nucleus metabolism, Cell Nucleus virology
- Abstract
Flaviviridae is a family of positive-stranded RNA viruses, including human pathogens, such as Japanese encephalitis virus (JEV), dengue virus (DENV), Zika virus (ZIKV), and West Nile virus (WNV). Nuclear localization of the viral core protein is conserved among Flaviviridae, and this feature may be targeted for developing broad-ranging anti-flavivirus drugs. However, the mechanism of core protein translocation to the nucleus and the importance of nuclear translocation in the viral life cycle remain unknown. We aimed to identify the molecular mechanism underlying core protein nuclear translocation. We identified importin-7 (IPO7), an importin-β family protein, as a nuclear carrier for Flaviviridae core proteins. Nuclear import assays revealed that core protein was transported into the nucleus via IPO7, whereas IPO7 deletion by CRISPR/Cas9 impaired their nuclear translocation. To understand the importance of core protein nuclear translocation, we evaluated the production of infectious virus or single-round-infectious-particles in wild-type or IPO7-deficient cells; both processes were significantly impaired in IPO7-deficient cells, whereas intracellular infectious virus levels were equivalent in wild-type and IPO7-deficient cells. These results suggest that IPO7-mediated nuclear translocation of core proteins is involved in the release of infectious virus particles of flaviviruses., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Itoh et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)
- Published
- 2024
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8. Hdm2 disrupts HdmX-mediated nuclear export of p53 by sequestering it in nucleus.
- Author
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Ni Y, Chen H, Cheng X, Sun B, Wu Z, Zhan Q, and Zhuang Z
- Subjects
- Humans, Nuclear Proteins metabolism, Nuclear Proteins genetics, Ubiquitination, Protein Binding, Tumor Suppressor Protein p53 metabolism, Tumor Suppressor Protein p53 genetics, Proto-Oncogene Proteins c-mdm2 metabolism, Active Transport, Cell Nucleus, Cell Nucleus metabolism, Proto-Oncogene Proteins metabolism, Cell Cycle Proteins metabolism, Cell Cycle Proteins genetics
- Abstract
Dysfunction of the tumor suppressor p53 occurs in most human cancers, Hdm2 and HdmX play critical roles in p53 inactivation and degradation. Under unstressed conditions, HdmX binds to p53 like Hdm2, but HdmX cannot directly induce p53 degradation. Moreover, HdmX has been reported to stimulate Hdm2-mediated ubiquitination and degradation of p53. Here we reported that HdmX promoted the nuclear export of p53 independent of Hdm2 in living cells using FRET technology. Whereas, Hdm2 impeded HdmX-mediated nuclear export of p53 by sequestering it in nucleus. Interestingly, the C-terminal RING domain mutant Hdm2
C464A formed heterooligomers with p53 in nucleus, which was inhibited by HdmX. The heterooligomers were located near PML-NBs. This study indicate that the nuclear Hdm2-HdmX interaction aborts the HdmX-mediated nuclear export of p53., Competing Interests: Declaration of competing interest The authors declare that they have no conflicts of interest with the contents of this article., (Copyright © 2024 Elsevier Inc. All rights reserved.)- Published
- 2024
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9. PDIA3 orchestrates effector T cell program by serving as a chaperone to facilitate the non-canonical nuclear import of STAT1 and PKM2.
- Author
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Yang CL, Wang FX, Luo JH, Rong SJ, Lu WY, Chen QJ, Xiao J, Wang T, Song DN, Liu J, Mo Q, Li S, Chen Y, Wang YN, Liu YJ, Yan T, Gu WK, Zhang S, Xiong F, Yu QL, Zhang ZY, Yang P, Liu SW, Eizirik D, Dong LL, Sun F, and Wang CY
- Subjects
- Humans, Mice, Animals, Membrane Proteins metabolism, Membrane Proteins genetics, Active Transport, Cell Nucleus, Carrier Proteins metabolism, Signal Transduction, Thyroid Hormone-Binding Proteins, NFATC Transcription Factors metabolism, Lymphocyte Activation, Thyroid Hormones metabolism, Gene Expression Regulation, Th17 Cells metabolism, Th17 Cells immunology, Th1 Cells immunology, Th1 Cells metabolism, Disease Models, Animal, Pyruvate Kinase, Protein Disulfide-Isomerases metabolism, Protein Disulfide-Isomerases genetics, Arthritis, Rheumatoid metabolism, STAT1 Transcription Factor metabolism
- Abstract
Dysregulated T cell activation underpins the immunopathology of rheumatoid arthritis (RA), yet the machineries that orchestrate T cell effector program remain incompletely understood. Herein, we leveraged bulk and single-cell RNA sequencing data from RA patients and validated protein disulfide isomerase family A member 3 (PDIA3) as a potential therapeutic target. PDIA3 is remarkably upregulated in pathogenic CD4 T cells derived from RA patients and positively correlates with C-reactive protein level and disease activity score 28. Pharmacological inhibition or genetic ablation of PDIA3 alleviates RA-associated articular pathology and autoimmune responses. Mechanistically, T cell receptor signaling triggers intracellular calcium flux to activate NFAT1, a process that is further potentiated by Wnt5a under RA settings. Activated NFAT1 then directly binds to the Pdia3 promoter to enhance the expression of PDIA3, which complexes with STAT1 or PKM2 to facilitate their nuclear import for transcribing T helper 1 (Th1) and Th17 lineage-related genes, respectively. This non-canonical regulatory mechanism likely occurs under pathological conditions, as PDIA3 could only be highly induced following aberrant external stimuli. Together, our data support that targeting PDIA3 is a vital strategy to mitigate autoimmune diseases, such as RA, in clinical settings., Competing Interests: Declaration of interests The authors declare that they have no conflict of interest., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2024
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10. Regulated circRNA nuclear export in neuronal differentiation.
- Author
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Li D and Huang Y
- Subjects
- Humans, Animals, Cell Nucleus metabolism, RNA, Circular metabolism, RNA, Circular genetics, Neurons metabolism, Neurons cytology, Active Transport, Cell Nucleus, Cell Differentiation, RNA metabolism
- Abstract
Multiple mechanisms have been reported for how circular RNAs (circRNAs) are exported to the cytoplasm. A recent paper by Cao et al. shows that export of a subset of circRNAs with (A)-rich motifs, including one with a clear function, is regulated during neuronal development via a novel mechanism., Competing Interests: Declaration of interests The authors have no conflicts of interests to declare., (Copyright © 2024 Elsevier Ltd. All rights reserved.)
- Published
- 2024
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11. Physiological regulation of neuronal Wnt activity is essential for TDP-43 localization and function.
- Author
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Zhang N, Westerhaus A, Wilson M, Wang E, Goff L, and Sockanathan S
- Subjects
- Humans, Animals, Mice, Amyotrophic Lateral Sclerosis metabolism, Amyotrophic Lateral Sclerosis genetics, Amyotrophic Lateral Sclerosis pathology, Active Transport, Cell Nucleus, Cell Nucleus metabolism, DNA-Binding Proteins metabolism, DNA-Binding Proteins genetics, Wnt Signaling Pathway, Neurons metabolism, Phosphoric Diester Hydrolases metabolism, Phosphoric Diester Hydrolases genetics
- Abstract
Nuclear exclusion of the RNA- and DNA-binding protein TDP-43 can induce neurodegeneration in different diseases. Diverse processes have been implicated to influence TDP-43 mislocalization, including disrupted nucleocytoplasmic transport (NCT); however, the physiological pathways that normally ensure TDP-43 nuclear localization are unclear. The six-transmembrane enzyme glycerophosphodiester phosphodiesterase 2 (GDE2 or GDPD5) cleaves the glycosylphosphatidylinositol (GPI) anchor that tethers some proteins to the membrane. Here we show that GDE2 maintains TDP-43 nuclear localization by regulating the dynamics of canonical Wnt signaling. Ablation of GDE2 causes aberrantly sustained Wnt activation in adult neurons, which is sufficient to cause NCT deficits, nuclear pore abnormalities, and TDP-43 nuclear exclusion. Disruption of GDE2 coincides with TDP-43 abnormalities in postmortem tissue from patients with amyotrophic lateral sclerosis (ALS). Further, GDE2 deficits are evident in human neural cell models of ALS, which display erroneous Wnt activation that, when inhibited, increases mRNA levels of genes regulated by TDP-43. Our study identifies GDE2 as a critical physiological regulator of Wnt signaling in adult neurons and highlights Wnt pathway activation as an unappreciated mechanism contributing to nucleocytoplasmic transport and TDP-43 abnormalities in disease., (© 2024. The Author(s).)
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- 2024
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12. Exportin 1 governs the immunosuppressive functions of myeloid-derived suppressor cells in tumors through ERK1/2 nuclear export.
- Author
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Daneshmandi S, Yan Q, Choi JE, Katsuta E, MacDonald CR, Goruganthu M, Roberts N, Repasky EA, Singh PK, Attwood K, Wang J, Landesman Y, McCarthy PL, and Mohammadpour H
- Subjects
- Animals, Humans, Mice, Mice, Inbred C57BL, Cell Differentiation, MAP Kinase Signaling System, Cell Line, Tumor, Interleukin-6 metabolism, Neoplasms immunology, Neoplasms pathology, Immune Tolerance, STAT3 Transcription Factor metabolism, Cell Nucleus metabolism, Exportin 1 Protein, Myeloid-Derived Suppressor Cells immunology, Myeloid-Derived Suppressor Cells metabolism, Receptors, Cytoplasmic and Nuclear metabolism, Karyopherins metabolism, Active Transport, Cell Nucleus
- Abstract
Myeloid-derived suppressor cells (MDSCs) are a main driver of immunosuppression in tumors. Understanding the mechanisms that determine the development and immunosuppressive function of these cells could provide new therapeutic targets to improve antitumor immunity. Here, using preclinical murine models, we discovered that exportin 1 (XPO1) expression is upregulated in tumor MDSCs and that this upregulation is induced by IL-6-induced STAT3 activation during MDSC differentiation. XPO1 blockade transforms MDSCs into T-cell-activating neutrophil-like cells, enhancing the antitumor immune response and restraining tumor growth. Mechanistically, XPO1 inhibition leads to the nuclear entrapment of ERK1/2, resulting in the prevention of ERK1/2 phosphorylation following the IL-6-mediated activation of the MAPK signaling pathway. Similarly, XPO1 blockade in human MDSCs induces the formation of neutrophil-like cells with immunostimulatory functions. Therefore, our findings revealed a critical role for XPO1 in MDSC differentiation and suppressive functions; exploiting these new discoveries revealed new targets for reprogramming immunosuppressive MDSCs to improve cancer therapeutic responses., (© 2024. The Author(s), under exclusive licence to CSI and USTC.)
- Published
- 2024
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13. Nanoassemblies designed for efficient nuclear targeting.
- Author
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Skowicki M, Tarvirdipour S, Kraus M, Schoenenberger CA, and Palivan CG
- Subjects
- Humans, Animals, Drug Delivery Systems, Nanoparticles chemistry, Peptides chemistry, Peptides administration & dosage, Polymers chemistry, Active Transport, Cell Nucleus, Drug Carriers chemistry, Nuclear Localization Signals, Cell Nucleus metabolism
- Abstract
One of the key aspects of coping efficiently with complex pathological conditions is delivering the desired therapeutic compounds with precision in both space and time. Therefore, the focus on nuclear-targeted delivery systems has emerged as a promising strategy with high potential, particularly in gene therapy and cancer treatment. Here, we explore the design of supramolecular nanoassemblies as vehicles to deliver specific compounds to the nucleus, with the special focus on polymer and peptide-based carriers that expose nuclear localization signals. Such nanoassemblies aim at maximizing the concentration of genetic and therapeutic agents within the nucleus, thereby optimizing treatment outcomes while minimizing off-target effects. A complex scenario of conditions, including cellular uptake, endosomal escape, and nuclear translocation, requires fine tuning of the nanocarriers' properties. First, we introduce the principles of nuclear import and the role of nuclear pore complexes that reveal strategies for targeting nanosystems to the nucleus. Then, we provide an overview of cargoes that rely on nuclear localization for optimal activity as their integrity and accumulation are crucial parameters to consider when designing a suitable delivery system. Considering that they are in their early stages of research, we present various cargo-loaded peptide- and polymer nanoassemblies that promote nuclear targeting, emphasizing their potential to enhance therapeutic response. Finally, we briefly discuss further advancements for more precise and effective nuclear delivery., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.)
- Published
- 2024
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14. Genome-wide quantification of RNA flow across subcellular compartments reveals determinants of the mammalian transcript life cycle.
- Author
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Ietswaart R, Smalec BM, Xu A, Choquet K, McShane E, Jowhar ZM, Guegler CK, Baxter-Koenigs AR, West ER, Fu BXH, Gilbert L, Floor SN, and Churchman LS
- Subjects
- Animals, Humans, Mice, Cytoplasm metabolism, Cytoplasm genetics, RNA Stability, Active Transport, Cell Nucleus, Polyribosomes metabolism, Polyribosomes genetics, Machine Learning, RNA-Binding Proteins metabolism, RNA-Binding Proteins genetics, Exosomes metabolism, Exosomes genetics, RNA, Messenger metabolism, RNA, Messenger genetics, Cell Nucleus metabolism, Cell Nucleus genetics, DEAD-box RNA Helicases metabolism, DEAD-box RNA Helicases genetics, Chromatin metabolism, Chromatin genetics
- Abstract
Dissecting the regulatory mechanisms controlling mammalian transcripts from production to degradation requires quantitative measurements of mRNA flow across the cell. We developed subcellular TimeLapse-seq to measure the rates at which RNAs are released from chromatin, exported from the nucleus, loaded onto polysomes, and degraded within the nucleus and cytoplasm in human and mouse cells. These rates varied substantially, yet transcripts from genes with related functions or targeted by the same transcription factors and RNA-binding proteins flowed across subcellular compartments with similar kinetics. Verifying these associations uncovered a link between DDX3X and nuclear export. For hundreds of RNA metabolism genes, most transcripts with retained introns were degraded by the nuclear exosome, while the remaining molecules were exported with stable cytoplasmic lifespans. Transcripts residing on chromatin for longer had extended poly(A) tails, whereas the reverse was observed for cytoplasmic mRNAs. Finally, machine learning identified molecular features that predicted the diverse life cycles of mRNAs., Competing Interests: Declaration of interests R.I. is a founder, board member, and/or shareholder of Cellforma, unrelated to the present work., (Copyright © 2024 Elsevier Inc. All rights reserved.)
- Published
- 2024
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15. AKT-dependent nuclear localization of EPRS1 activates PARP1 in breast cancer cells.
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Zin I, China A, Khan K, Nag JK, Vasu K, Deshpande GM, Ghosh PK, Khan D, Ramachandiran I, Ganguly S, Tamagno I, Willard B, Gogonea V, and Fox PL
- Subjects
- Humans, Female, Cell Line, Tumor, PTEN Phosphohydrolase metabolism, PTEN Phosphohydrolase genetics, Amino Acyl-tRNA Synthetases metabolism, Amino Acyl-tRNA Synthetases genetics, Active Transport, Cell Nucleus, Nuclear Localization Signals metabolism, Breast Neoplasms metabolism, Breast Neoplasms genetics, Breast Neoplasms pathology, Poly (ADP-Ribose) Polymerase-1 metabolism, Poly (ADP-Ribose) Polymerase-1 genetics, Cell Nucleus metabolism, Proto-Oncogene Proteins c-akt metabolism, Proto-Oncogene Proteins c-akt genetics
- Abstract
Glutamyl-prolyl-tRNA synthetase (EPRS1) is a bifunctional aminoacyl-tRNA-synthetase (aaRS) essential for decoding the genetic code. EPRS1 resides, with seven other aaRSs and three noncatalytic proteins, in the cytoplasmic multi-tRNA synthetase complex (MSC). Multiple MSC-resident aaRSs, including EPRS1, exhibit stimulus-dependent release from the MSC to perform noncanonical activities distinct from their primary function in protein synthesis. Here, we show EPRS1 is present in both cytoplasm and nucleus of breast cancer cells with constitutively low phosphatase and tensin homolog (PTEN) expression. EPRS1 is primarily cytosolic in PTEN-expressing cells, but chemical or genetic inhibition of PTEN, or chemical or stress-mediated activation of its target, AKT, induces EPRS1 nuclear localization. Likewise, preferential nuclear localization of EPRS1 was observed in invasive ductal carcinoma that were also P-Ser
473 -AKT+ . EPRS1 nuclear transport requires a nuclear localization signal (NLS) within the linker region that joins the catalytic glutamyl-tRNA synthetase and prolyl-tRNA synthetase domains. Nuclear EPRS1 interacts with poly(ADP-ribose) polymerase 1 (PARP1), a DNA-damage sensor that directs poly(ADP-ribosyl)ation (PARylation) of proteins. EPRS1 is a critical regulator of PARP1 activity as shown by markedly reduced ADP-ribosylation in EPRS1 knockdown cells. Moreover, EPRS1 and PARP1 knockdown comparably alter the expression of multiple tumor-related genes, inhibit DNA-damage repair, reduce tumor cell survival, and diminish tumor sphere formation by breast cancer cells. EPRS1-mediated regulation of PARP1 activity provides a mechanistic link between PTEN loss in breast cancer cells, PARP1 activation, and cell survival and tumor growth. Targeting the noncanonical activity of EPRS1, without inhibiting canonical tRNA ligase activity, provides a therapeutic approach potentially supplementing existing PARP1 inhibitors., Competing Interests: Competing interests statement:The authors declare no competing interest.- Published
- 2024
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16. Increased level of TXNIP and nuclear translocation of TXN is associated with end stage renal disease and development of multiplex renal tumours.
- Author
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Beothe T, Docs J, Kovacs G, and Peterfi L
- Subjects
- Humans, Male, Middle Aged, Female, Carcinoma, Renal Cell metabolism, Carcinoma, Renal Cell pathology, Aged, Cell Nucleus metabolism, Adult, Active Transport, Cell Nucleus, Kidney Neoplasms metabolism, Kidney Neoplasms pathology, Carrier Proteins metabolism, Thioredoxins metabolism, Kidney Failure, Chronic metabolism
- Abstract
Background: End-stage and acquired cystic renal disease (ESRD/ACRD) kidneys are characterized by inflammatory remodelling and multiplex renal cell carcinomas (RCC). Eosinophilic vacuolated tumour (EVT) occurs exclusively in ACRD. The aim of this study was to identify the involvement of thioredoxin-interacting protein (TXNIP) and thioredoxin (TXN) in ESRD/ACRD pathology., Methods: Expression of TXNIP and TXN was examined in histological slides of 6 ESRD and 6 ACRD kidneys, precursor lesions and associated tumours as well as of RCCs from the general population by immunohistochemistry., Results: Strong TXNIP expression was seen in epithelial cells, myo-fibroblasts and endothelial cells and weak TXN expression in ESRD/ACRD kidneys and tumours. In ACRD specific EVT and its precursors TXN were translocated into nuclei., Conclusion: The impaired TXNIP/TXN redox homeostasis might be associated with development of multiplex cancer especially of EVT in ESRD/ACRD kidney., (© 2024. The Author(s).)
- Published
- 2024
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17. Functional Analysis of GRSF1 in the Nuclear Export and Translation of Influenza A Virus mRNAs.
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Schmierer J and Takimoto T
- Subjects
- Humans, Animals, Influenza A virus genetics, Influenza A virus physiology, Influenza A virus metabolism, Virus Replication, Influenza A Virus, H1N1 Subtype genetics, Influenza A Virus, H1N1 Subtype metabolism, Influenza A Virus, H1N1 Subtype physiology, Cell Nucleus metabolism, Cell Nucleus virology, 5' Untranslated Regions genetics, Nucleocapsid Proteins metabolism, Nucleocapsid Proteins genetics, Madin Darby Canine Kidney Cells, HEK293 Cells, RNA-Binding Proteins metabolism, RNA-Binding Proteins genetics, Dogs, Influenza, Human virology, Influenza, Human metabolism, Influenza, Human genetics, Mutation, Host-Pathogen Interactions genetics, Viral Core Proteins metabolism, Viral Core Proteins genetics, Active Transport, Cell Nucleus, RNA, Messenger genetics, RNA, Messenger metabolism, Protein Biosynthesis, RNA, Viral genetics, RNA, Viral metabolism
- Abstract
Influenza A viruses (IAV) utilize host proteins throughout their life cycle to infect and replicate in their hosts. We previously showed that host adaptive mutations in avian IAV PA help recruit host protein G-Rich RNA Sequence Binding Factor 1 (GRSF1) to the nucleoprotein (NP) 5' untranslated region (UTR), leading to the enhanced nuclear export and translation of NP mRNA. In this study, we evaluated the impact of GRSF1 in the viral life cycle. We rescued and characterized a 2009 pH1N1 virus with a mutated GRSF1 binding site in the 5' UTR of NP mRNA. Mutant viral growth was attenuated relative to pH1N1 wild-type (WT) in mammalian cells. We observed a specific reduction in the NP protein production and cytosolic accumulation of NP mRNAs, indicating a critical role of GRSF1 in the nuclear export of IAV NP mRNAs. Further, in vitro-transcribed mutated NP mRNA was translated less efficiently than WT NP mRNA in transfected cells. Together, these findings show that GRSF1 binding is important for both mRNA nuclear export and translation and affects overall IAV growth. Enhanced association of GRSF1 to NP mRNA by PA mutations leads to rapid virus growth, which could be a key process of mammalian host adaptation of IAV.
- Published
- 2024
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18. RNF213 promotes Treg cell differentiation by facilitating K63-linked ubiquitination and nuclear translocation of FOXO1.
- Author
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Yang X, Zhu X, Sheng J, Fu Y, Nie D, You X, Chen Y, Yang X, Ling Q, Zhang H, Li X, and Hu S
- Subjects
- Animals, Mice, Humans, Mice, Inbred C57BL, Cell Nucleus metabolism, Multiple Sclerosis immunology, Multiple Sclerosis metabolism, Multiple Sclerosis genetics, Multiple Sclerosis pathology, Active Transport, Cell Nucleus, Female, Mice, Knockout, Encephalomyelitis, Autoimmune, Experimental immunology, Encephalomyelitis, Autoimmune, Experimental metabolism, Encephalomyelitis, Autoimmune, Experimental genetics, HEK293 Cells, Ubiquitination, T-Lymphocytes, Regulatory immunology, T-Lymphocytes, Regulatory metabolism, Ubiquitin-Protein Ligases metabolism, Ubiquitin-Protein Ligases genetics, Cell Differentiation, Forkhead Box Protein O1 metabolism, Forkhead Box Protein O1 genetics, Interferon-beta metabolism
- Abstract
Autoreactive CD4
+ T helper cells are critical players that orchestrate the immune response both in multiple sclerosis (MS) and in other neuroinflammatory autoimmune diseases. Ubiquitination is a posttranslational protein modification involved in regulating a variety of cellular processes, including CD4+ T cell differentiation and function. However, only a limited number of E3 ubiquitin ligases have been characterized in terms of their biological functions, particularly in CD4+ T cell differentiation and function. In this study, we found that the RING finger protein 213 (RNF213) specifically promoted regulatory T (Treg) cell differentiation in CD4+ T cells and attenuated autoimmune disease development in an FOXO1-dependent manner. Mechanistically, RNF213 interacts with Forkhead Box Protein O1 (FOXO1) and promotes nuclear translocation of FOXO1 by K63-linked ubiquitination. Notably, RNF213 expression in CD4+ T cells was induced by IFN-β and exerts a crucial role in the therapeutic efficacy of IFN-β for MS. Together, our study findings collectively emphasize the pivotal role of RNF213 in modulating adaptive immune responses. RNF213 holds potential as a promising therapeutic target for addressing disorders associated with Treg cells., (© 2024. The Author(s).)- Published
- 2024
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19. O-GlcNAcylated RALY Contributes to Hepatocellular Carcinoma Cells Proliferation by Regulating USP22 mRNA Nuclear Export.
- Author
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Liu S, Lv Q, Mao X, Dong H, Xu W, Du X, Jia W, Feng K, Zhang J, and Zhang Y
- Subjects
- Humans, Cell Line, Tumor, Animals, RNA, Messenger metabolism, RNA, Messenger genetics, Mice, Mice, Nude, Ubiquitination, Active Transport, Cell Nucleus, Carcinoma, Hepatocellular metabolism, Carcinoma, Hepatocellular pathology, Carcinoma, Hepatocellular genetics, Liver Neoplasms metabolism, Liver Neoplasms pathology, Liver Neoplasms genetics, Cell Proliferation, Ubiquitin Thiolesterase metabolism, Ubiquitin Thiolesterase genetics
- Abstract
Hepatocellular carcinoma (HCC) is one of the most prevalent and deadly tumors; however, its pathogenic mechanism remains largely elusive. In-depth researches are needed to reveal the expression regulatory mechanisms and functions of the RNA-binding protein RALY in HCC. Here, we identify RALY as a highly expressed oncogenic factor that affects HCC cells proliferation both in vitro and in vivo . O-GlcNAcylation of RALY at Ser176 enhances its stability by protecting RALY from TRIM27-mediated ubiquitination, thus maintaining hyper-expression of the RALY protein. Mechanistically, RALY interacts with USP22 messenger RNA, as revealed by RNA immunoprecipitation, to increase their cytoplasmic localization and protein expression, thereby promoting the proliferation of HCC cells. Furthermore, we develop a novel RALY protein degrader based on peptide proteolysis-targeting chimeras, named RALY-PROTAC, which we chemically synthesize by linking a RALY-targeting peptide with the E3 ubiquitin ligase recruitment ligand pomalidomide. In conclusion, our findings demonstrate a novel mechanism by which O-GlcNAcylation/RALY/USP22 mRNA axis aggravates HCC cells proliferation. RALY-PROTACs as degraders of the RALY protein exhibit potential as therapeutic drugs for RALY-overexpressing HCC., Competing Interests: Competing Interests: The authors have declared that no competing interest exists., (© The author(s).)
- Published
- 2024
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- View/download PDF
20. Nucleoporin Nup98 is an essential factor for ipo4 dependent protein import.
- Author
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Guo Y, Tao T, Wu T, Hou J, and Lin W
- Subjects
- Humans, Cell Nucleus metabolism, HeLa Cells, Protein Binding, Active Transport, Cell Nucleus, beta Karyopherins metabolism, beta Karyopherins genetics, Nuclear Pore Complex Proteins metabolism, Nuclear Pore Complex Proteins genetics
- Abstract
Nucleocytoplasmic transport of macromolecules is essential in eukaryotic cells. In this process, the karyopherins play a central role when they transport cargoes across the nuclear pore complex. Importin 4 belongs to the karyopherin β family. Many studies have focused on finding substrates for importin 4, but no direct mechanism studies of its precise transport function have been reported. Therefore, this paper mainly aimed to study the mechanism of nucleoporins in mediating nuclear import and export of importin 4. To address this question, we constructed shRNAs targeting Nup358, Nup153, Nup98, and Nup50. We found that depletion of Nup98 resulted in a shift in the subcellular localization of importin 4 from the cytoplasm to the nucleus. Mutational analysis demonstrated that Nup98 physically and functionally interacts with importin 4 through its N-terminal phenylalanine-glycine (FG) repeat region. Mutation of nine of these FG motifs to SG motifs significantly attenuated the binding of Nup98 to importin 4, and we further confirmed the essential role of the six FG motifs in amino acids 121-360 of Nup98 in binding with importin 4. In vitro transport assay also confirmed that VDR, the substrate of importin 4, could not be transported into the nucleus after Nup98 knockdown. Overall, our results showed that Nup98 is required for efficient importin 4-mediated transport. This is the first study to reveal the mechanism of importin 4 in transporting substrates into the nucleus., (© 2024 Wiley Periodicals LLC.)
- Published
- 2024
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- View/download PDF
21. UBAP2L ensures homeostasis of nuclear pore complexes at the intact nuclear envelope.
- Author
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Liao Y, Andronov L, Liu X, Lin J, Guerber L, Lu L, Agote-Arán A, Pangou E, Ran L, Kleiss C, Qu M, Schmucker S, Cirillo L, Zhang Z, Riveline D, Gotta M, Klaholz BP, and Sumara I
- Subjects
- Humans, Active Transport, Cell Nucleus, HeLa Cells, Homeostasis, Membrane Glycoproteins, Nuclear Pore Complex Proteins metabolism, Nuclear Envelope metabolism, Nuclear Pore metabolism, Carrier Proteins metabolism
- Abstract
Assembly of macromolecular complexes at correct cellular sites is crucial for cell function. Nuclear pore complexes (NPCs) are large cylindrical assemblies with eightfold rotational symmetry, built through hierarchical binding of nucleoporins (Nups) forming distinct subcomplexes. Here, we uncover a role of ubiquitin-associated protein 2-like (UBAP2L) in the assembly and stability of properly organized and functional NPCs at the intact nuclear envelope (NE) in human cells. UBAP2L localizes to the nuclear pores and facilitates the formation of the Y-complex, an essential scaffold component of the NPC, and its localization to the NE. UBAP2L promotes the interaction of the Y-complex with POM121 and Nup153, the critical upstream factors in a well-defined sequential order of Nups assembly onto NE during interphase. Timely localization of the cytoplasmic Nup transport factor fragile X-related protein 1 (FXR1) to the NE and its interaction with the Y-complex are likewise dependent on UBAP2L. Thus, this NPC biogenesis mechanism integrates the cytoplasmic and the nuclear NPC assembly signals and ensures efficient nuclear transport, adaptation to nutrient stress, and cellular proliferative capacity, highlighting the importance of NPC homeostasis at the intact NE., (© 2024 Liao et al.)
- Published
- 2024
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22. dsRNA formation leads to preferential nuclear export and gene expression.
- Author
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Coban I, Lamping JP, Hirsch AG, Wasilewski S, Shomroni O, Giesbrecht O, Salinas G, and Krebber H
- Subjects
- Cytoplasm metabolism, DEAD-box RNA Helicases metabolism, DEAD-box RNA Helicases genetics, Nucleocytoplasmic Transport Proteins metabolism, Nucleocytoplasmic Transport Proteins genetics, RNA-Binding Proteins metabolism, RNA-Binding Proteins genetics, Saccharomyces cerevisiae Proteins metabolism, Saccharomyces cerevisiae Proteins genetics, Active Transport, Cell Nucleus, Cell Nucleus metabolism, Gene Expression Regulation, Fungal, RNA Transport, RNA, Antisense metabolism, RNA, Antisense genetics, RNA, Double-Stranded metabolism, RNA, Double-Stranded genetics, RNA, Messenger metabolism, RNA, Messenger genetics, Saccharomyces cerevisiae genetics, Saccharomyces cerevisiae metabolism
- Abstract
When mRNAs have been transcribed and processed in the nucleus, they are exported to the cytoplasm for translation. This export is mediated by the export receptor heterodimer Mex67-Mtr2 in the yeast Saccharomyces cerevisiae (TAP-p15 in humans)
1,2 . Interestingly, many long non-coding RNAs (lncRNAs) also leave the nucleus but it is currently unclear why they move to the cytoplasm3 . Here we show that antisense RNAs (asRNAs) accelerate mRNA export by annealing with their sense counterparts through the helicase Dbp2. These double-stranded RNAs (dsRNAs) dominate export compared with single-stranded RNAs (ssRNAs) because they have a higher capacity and affinity for the export receptor Mex67. In this way, asRNAs boost gene expression, which is beneficial for cells. This is particularly important when the expression program changes. Consequently, the degradation of dsRNA, or the prevention of its formation, is toxic for cells. This mechanism illuminates the general cellular occurrence of asRNAs and explains their nuclear export., (© 2024. The Author(s).)- Published
- 2024
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23. Nucleocytoplasmic transport rates are regulated by cellular processes that modulate GTP availability.
- Author
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Scott KL, Halfmann CT, Hoefakker AD, Purkayastha P, Wang TC, Lele TP, and Roux KJ
- Subjects
- Humans, Cell Nucleus metabolism, Cell Movement, Nuclear Pore metabolism, Nuclear Pore genetics, Animals, Nuclear Envelope metabolism, Cytoskeleton metabolism, Protein Biosynthesis, Cytoplasm metabolism, Guanosine Triphosphate metabolism, Active Transport, Cell Nucleus, ran GTP-Binding Protein metabolism, ran GTP-Binding Protein genetics
- Abstract
Nucleocytoplasmic transport (NCT), the facilitated diffusion of cargo molecules between the nucleus and cytoplasm through nuclear pore complexes (NPCs), enables numerous fundamental eukaryotic cellular processes. Ran GTPase uses cellular energy in the direct form of GTP to create a gradient across the nuclear envelope (NE) that drives the majority of NCT. We report here that changes in GTP availability resulting from altered cellular physiology modulate the rate of NCT, as monitored using synthetic and natural cargo, and the dynamics of Ran itself. Cell migration, cell spreading, and/or modulation of the cytoskeleton or its connection to the nucleus alter GTP availability and thus rates of NCT, regulating RNA export and protein synthesis. These findings support a model in which changes in cellular physiology that alter GTP availability can regulate the rate of NCT, impacting fundamental cellular processes that extensively utilize NCT., (© 2024 Scott et al.)
- Published
- 2024
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- View/download PDF
24. Nuclear transport receptors underpin plastidial retrograde signaling.
- Author
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Jia M and Gu Y
- Subjects
- Plastids metabolism, Active Transport, Cell Nucleus, Signal Transduction
- Published
- 2024
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25. The varicella-zoster virus ORF16 protein promotes both the nuclear transport and the protein abundance of the viral DNA polymerase subunit ORF28.
- Author
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Lin HS, Li CH, Chen LW, Wang SS, Chen LY, Hung CH, Lin CL, and Chang PJ
- Subjects
- Humans, Cell Nucleus metabolism, Cell Nucleus virology, Cytoplasm metabolism, Cytoplasm virology, Cell Line, DNA Replication, Herpesvirus 3, Human genetics, Herpesvirus 3, Human metabolism, Viral Proteins metabolism, Viral Proteins genetics, Virus Replication, Active Transport, Cell Nucleus, DNA-Directed DNA Polymerase metabolism, DNA-Directed DNA Polymerase genetics
- Abstract
Although all herpesviruses utilize a highly conserved replication machinery to amplify their viral genomes, different members may have unique strategies to modulate the assembly of their replication components. Herein, we characterize the subcellular localization of seven essential replication proteins of varicella-zoster virus (VZV) and show that several viral replication enzymes such as the DNA polymerase subunit ORF28, when expressed alone, are localized in the cytoplasm. The nuclear import of ORF28 can be mediated by the viral DNA polymerase processivity factor ORF16. Besides, ORF16 could markedly enhance the protein abundance of ORF28. Noteworthily, an ORF16 mutant that is defective in nuclear transport still retained the ability to enhance ORF28 abundance. The low abundance of ORF28 in transfected cells was due to its rapid degradation mediated by the ubiquitin-proteasome system. We additionally reveal that radicicol, an inhibitor of the chaperone Hsp90, could disrupt the interaction between ORF16 and ORF28, thereby affecting the nuclear entry and protein abundance of ORF28. Collectively, our findings imply that the cytoplasmic retention and rapid degradation of ORF28 may be a key regulatory mechanism for VZV to prevent untimely viral DNA replication, and suggest that Hsp90 is required for the interaction between ORF16 and ORF28., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)
- Published
- 2024
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- View/download PDF
26. Significance of signal recognition particle 9 nuclear translocation: Implications for pancreatic cancer prognosis and functionality.
- Author
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Sato H, Meng S, Sasaki K, Kobayashi S, Kido K, Tsuji Y, Arao Y, Saito Y, Iwagami Y, Yamada D, Tomimaru Y, Noda T, Takahashi H, Motooka D, Uchida S, Ofusa K, Satoh T, Doki Y, Eguchi H, Hara T, and Ishii H
- Subjects
- Humans, Prognosis, Male, Female, Middle Aged, Aged, Cell Line, Tumor, Signal Recognition Particle metabolism, Signal Recognition Particle genetics, Active Transport, Cell Nucleus, Serine-Arginine Splicing Factors metabolism, Serine-Arginine Splicing Factors genetics, Adult, Gene Expression Regulation, Neoplastic, Pancreatic Neoplasms pathology, Pancreatic Neoplasms genetics, Pancreatic Neoplasms metabolism, Pancreatic Neoplasms mortality, Cell Nucleus metabolism
- Abstract
Signal recognition particles (SRPs) are essential for regulating intracellular protein transport and secretion. Patients with tumors with high SRP9 expression tend to have a poorer overall survival. However, to the best of our knowledge, no reports have described the relationship between SRP9 localization and prognosis in pancreatic cancer. Thus, the present study aimed to investigate this relationship. Immunohistochemical staining for SRP9 using excised specimens from pancreatic cancer surgery cases without preoperative chemotherapy or radiotherapy showed that SRP9 was preferentially expressed in the nucleus of the cancerous regions in some cases, which was hardly detected in other cases, indicating that SRP9 was transported to the nucleus in the former cases. To compare the prognosis of patients with SRP9 nuclear translocation, patients were divided into two groups: Those with a nuclear translocation rate of >50% and those with a nuclear translocation rate of ≤50%. The nuclear translocation rate of >50% group had a significantly better recurrence‑free survival than the nuclear translocation rate of ≤50% group (P=0.037). Subsequent in vitro experiments were conducted; notably, the nuclear translocation rate of SRP9 was reduced under amino acid‑deficient conditions, suggesting that multiple factors are involved in this phenomenon. To further study the function of SRP9 nuclear translocation, in vitro experiments were performed by introducing SRP9 splicing variants (v1 and v2) and their deletion mutants lacking C‑terminal regions into MiaPaCa pancreatic cancer cells. The results demonstrated that both splicing variants showed nuclear translocation regardless of the C‑terminal deletions, suggesting the role of the N‑terminal regions. Given that SRP9 is an RNA‑binding protein, the study of RNA immunoprecipitation revealed that signaling pathways involved in cancer progression and protein translation were downregulated in nuclear‑translocated v1 and v2. Undoubtedly, further studies of the nuclear translocation of SRP9 will open an avenue to optimize the precise evaluation and therapeutic control of pancreatic cancer.
- Published
- 2024
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- View/download PDF
27. The bioenergetics of nucleocytoplasmic transport.
- Author
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Luxton GWG
- Subjects
- Humans, Cell Movement, Cell Nucleus metabolism, Guanosine Triphosphate metabolism, Protein Biosynthesis, Active Transport, Cell Nucleus, Energy Metabolism
- Abstract
How nucleocytoplasmic transport (NCT) rates change due to cellular physiology-mediated fluctuations in GTP availability remains unclear. In this issue, Scott et al. (https://doi.org/10.1083/jcb.202308152) demonstrate that cell migration, spreading, and nucleocytoskeletal coupling impact GTP levels, thereby regulating NCT, RNA export, and protein synthesis., (© 2024 Luxton.)
- Published
- 2024
- Full Text
- View/download PDF
28. The PB2 I714S mutation influenced mammalian adaptation of the H3N2 canine influenza virus by interfering with nuclear import efficiency and RNP complex assembly.
- Author
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Li X, Jia T, Wang K, Wang L, Zhou L, Li M, Zhu W, Shu Y, and Chen Y
- Subjects
- Animals, Dogs, Mice, Humans, Active Transport, Cell Nucleus, Virus Replication, Mutation, Madin Darby Canine Kidney Cells, Dog Diseases virology, Mice, Inbred BALB C, HEK293 Cells, Reassortant Viruses genetics, Influenza A Virus, H3N2 Subtype genetics, Influenza A Virus, H3N2 Subtype physiology, Viral Proteins genetics, Viral Proteins metabolism, RNA-Dependent RNA Polymerase genetics, RNA-Dependent RNA Polymerase metabolism, Orthomyxoviridae Infections virology, Ribonucleoproteins genetics, Ribonucleoproteins metabolism
- Abstract
Avian influenza viruses (AIVs) are the origin of multiple mammal influenza viruses. The genetic determinants of AIVs adapted to humans have been widely elucidated, however, the molecular mechanism of cross-species transmission and adaptation of AIVs to canines are still poorly understood. In this study, two H3N2 influenza viruses isolated from a live poultry market (A/environment/Guangxi/13431/2018, GX13431) and a swab sample from a canine (A/canine/Guangdong/0601/2019, GD0601) were used to investigate the possible molecular basis that determined H3N2 AIV adapting to canine. We found that GD0601 exhibited more robust polymerase activity in cells and higher pathogenicity in mice compared with its evolution ancestor H3N2 AIV GX13431. A series of reassortments of the ribonucleoprotein (RNP) complex showed that the PB2 subunit was the crucial factor that conferred high polymerase activity of GD0601, and the substitution of I714S in the PB2 subunit of GD0601 attenuated the replication and pathogenicity in mammal cells and the mouse model. Mechanistically, the reverse mutation of I714S in the PB2 polymerase subunit which was identified in AIV GX13431 reduced the nuclear import efficiency of PB2 protein and interfered with the interactions of PB2-PA/NP that affected the assembly of the viral RNP complex. Our study reveals amino acid mutation at the position of 714 in the nuclear localization signal (NLS) area in PB2 plays an important role in overcoming the barrier from poultry to mammals of the H3N2 canine influenza virus and provides clues for further study of mammalian adaptation mechanism of AIVs.
- Published
- 2024
- Full Text
- View/download PDF
29. Cytoplasmic nucleoporin assemblage: the cellular artwork in physiology and disease.
- Author
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Lin J and Sumara I
- Subjects
- Humans, Animals, Nuclear Pore metabolism, Active Transport, Cell Nucleus, Nuclear Pore Complex Proteins metabolism, Cytoplasm metabolism
- Abstract
Nucleoporins, essential proteins building the nuclear pore, are pivotal for ensuring nucleocytoplasmic transport. While traditionally confined to the nuclear envelope, emerging evidence indicates their presence in various cytoplasmic structures, suggesting potential non-transport-related roles. This review consolidates findings on cytoplasmic nucleoporin assemblies across different states, including normal physiological conditions, stress, and pathology, exploring their structural organization, formation dynamics, and functional implications. We summarize the current knowledge and the latest concepts on the regulation of nucleoporin homeostasis, aiming to enhance our understanding of their unexpected roles in physiological and pathological processes.
- Published
- 2024
- Full Text
- View/download PDF
30. Inhibition of RAN attenuates influenza a virus replication and nucleoprotein nuclear export.
- Author
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Cao L, She Z, Zhao Y, Cheng C, Li Y, Xu T, Mao H, Zhang Y, Hui X, Lin X, Wang T, Sun X, Huang K, Zhao L, and Jin M
- Subjects
- Humans, Animals, Dogs, Receptors, Cytoplasmic and Nuclear metabolism, Receptors, Cytoplasmic and Nuclear genetics, Madin Darby Canine Kidney Cells, Viral Nonstructural Proteins metabolism, Viral Nonstructural Proteins genetics, Mice, Piperidines pharmacology, Influenza, Human virology, A549 Cells, Nucleoproteins metabolism, Nucleoproteins genetics, HEK293 Cells, Cell Line, Cell Nucleus metabolism, Ribonucleoproteins metabolism, Ribonucleoproteins genetics, Virus Replication drug effects, ran GTP-Binding Protein metabolism, ran GTP-Binding Protein genetics, Antiviral Agents pharmacology, Influenza A virus drug effects, Influenza A virus physiology, Active Transport, Cell Nucleus, Exportin 1 Protein, Karyopherins metabolism, Karyopherins antagonists & inhibitors
- Abstract
Nuclear export of the viral ribonucleoprotein (vRNP) is a critical step in the influenza A virus (IAV) life cycle and may be an effective target for the development of anti-IAV drugs. The host factor ras-related nuclear protein (RAN) is known to participate in the life cycle of several viruses, but its role in influenza virus replication remains unknown. In the present study, we aimed to determine the function of RAN in influenza virus replication using different cell lines and subtype strains. We found that RAN is essential for the nuclear export of vRNP, as it enhances the binding affinity of XPO1 toward the viral nuclear export protein NS2. Depletion of RAN constrained the vRNP complex in the nucleus and attenuated the replication of various subtypes of influenza virus. Using in silico compound screening, we identified that bepotastine could dissociate the RAN-XPO1-vRNP trimeric complex and exhibit potent antiviral activity against influenza virus both in vitro and in vivo . This study demonstrates the important role of RAN in IAV replication and suggests its potential use as an antiviral target.
- Published
- 2024
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- View/download PDF
31. Pre-ribosomal particles from nucleoli to cytoplasm.
- Author
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Kubitscheck U and Siebrasse JP
- Subjects
- Humans, Animals, Ribosomes metabolism, Cell Nucleus metabolism, Cell Nucleolus metabolism, Nuclear Pore metabolism, Cytoplasm metabolism, Active Transport, Cell Nucleus
- Abstract
The analysis of nucleocytoplasmic transport of proteins and messenger RNA has been the focus of advanced microscopic approaches. Recently, it has been possible to identify and visualize individual pre-ribosomal particles on their way through the nuclear pore complex using both electron and light microscopy. In this review, we focused on the transport of pre-ribosomal particles in the nucleus on their way to and through the pores.
- Published
- 2024
- Full Text
- View/download PDF
32. Sculpting nuclear envelope identity from the endoplasmic reticulum during the cell cycle.
- Author
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Deolal P, Scholz J, Ren K, Bragulat-Teixidor H, and Otsuka S
- Subjects
- Endoplasmic Reticulum metabolism, Mitosis, Active Transport, Cell Nucleus, Nuclear Envelope metabolism, Nuclear Pore metabolism
- Abstract
The nuclear envelope (NE) regulates nuclear functions, including transcription, nucleocytoplasmic transport, and protein quality control. While the outer membrane of the NE is directly continuous with the endoplasmic reticulum (ER), the NE has an overall distinct protein composition from the ER, which is crucial for its functions. During open mitosis in higher eukaryotes, the NE disassembles during mitotic entry and then reforms as a functional territory at the end of mitosis to reestablish nucleocytoplasmic compartmentalization. In this review, we examine the known mechanisms by which the functional NE reconstitutes from the mitotic ER in the continuous ER-NE endomembrane system during open mitosis. Furthermore, based on recent findings indicating that the NE possesses unique lipid metabolism and quality control mechanisms distinct from those of the ER, we explore the maintenance of NE identity and homeostasis during interphase. We also highlight the potential significance of membrane junctions between the ER and NE.
- Published
- 2024
- Full Text
- View/download PDF
33. Not just binary: embracing the complexity of nuclear division dynamics.
- Author
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Walsh ME, King GA, and Ünal E
- Subjects
- Humans, Animals, Cell Nucleus metabolism, Nuclear Envelope metabolism, Chromosomes metabolism, Active Transport, Cell Nucleus, Cell Nucleus Division
- Abstract
Cell division presents a challenge for eukaryotic cells: how can chromosomes effectively segregate within the confines of a membranous nuclear compartment? Different organisms have evolved diverse solutions by modulating the degree of nuclear compartmentalization, ranging from complete nuclear envelope breakdown to complete maintenance of nuclear compartmentalization via nuclear envelope expansion. Many intermediate forms exist between these extremes, suggesting that nuclear dynamics during cell division are surprisingly plastic. In this review, we highlight the evolutionary diversity of nuclear divisions, focusing on two defining characteristics: (1) chromosome compartmentalization and (2) nucleocytoplasmic transport. Further, we highlight recent evidence that nuclear behavior during division can vary within different cellular contexts in the same organism. The variation observed within and between organisms underscores the dynamic evolution of nuclear divisions tailored to specific contexts and cellular requirements. In-depth investigation of diverse nuclear divisions will enhance our understanding of the nucleus, both in physiological and pathological states.
- Published
- 2024
- Full Text
- View/download PDF
34. Nesprin-2 coordinates opposing microtubule motors during nuclear migration in neurons.
- Author
-
Zhou C, Wu YK, Ishidate F, Fujiwara TK, and Kengaku M
- Subjects
- Animals, Mice, Active Transport, Cell Nucleus, Dynactin Complex metabolism, Dynactin Complex genetics, Cell Movement, Microfilament Proteins metabolism, Microfilament Proteins genetics, Nuclear Proteins metabolism, Nuclear Proteins genetics, Cerebellum metabolism, Cerebellum cytology, Binding Sites, Humans, Microtubules metabolism, Neurons metabolism, Kinesins metabolism, Kinesins genetics, Nerve Tissue Proteins metabolism, Nerve Tissue Proteins genetics, Dyneins metabolism, Cell Nucleus metabolism, Microtubule-Associated Proteins metabolism, Microtubule-Associated Proteins genetics
- Abstract
Nuclear migration is critical for the proper positioning of neurons in the developing brain. It is known that bidirectional microtubule motors are required for nuclear transport, yet the mechanism of the coordination of opposing motors is still under debate. Using mouse cerebellar granule cells, we demonstrate that Nesprin-2 serves as a nucleus-motor adaptor, coordinating the interplay of kinesin-1 and dynein. Nesprin-2 recruits dynein-dynactin-BicD2 independently of the nearby kinesin-binding LEWD motif. Both motor binding sites are required to rescue nuclear migration defects caused by the loss of function of Nesprin-2. In an intracellular cargo transport assay, the Nesprin-2 fragment encompassing the motor binding sites generates persistent movements toward both microtubule minus and plus ends. Nesprin-2 drives bidirectional cargo movements over a prolonged period along perinuclear microtubules, which advance during the migration of neurons. We propose that Nesprin-2 keeps the nucleus mobile by coordinating opposing motors, enabling continuous nuclear transport along advancing microtubules in migrating cells., (© 2024 Zhou et al.)
- Published
- 2024
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- View/download PDF
35. Exportin XPO6 upregulation activates the TLR2/MyD88/NF-κB signaling by facilitating TLR2 mRNA nuclear export in COPD pulmonary monocytes.
- Author
-
Wu Y, Gou Y, Wang T, Li P, Li Y, Lu X, Li W, and Liu Z
- Subjects
- Animals, Humans, Mice, Male, Active Transport, Cell Nucleus, Lung pathology, Lung immunology, Lung metabolism, Disease Models, Animal, Female, Toll-Like Receptor 2 metabolism, Toll-Like Receptor 2 genetics, Pulmonary Disease, Chronic Obstructive metabolism, Pulmonary Disease, Chronic Obstructive immunology, Myeloid Differentiation Factor 88 metabolism, Monocytes metabolism, Monocytes immunology, Monocytes drug effects, NF-kappa B metabolism, Signal Transduction, Up-Regulation, Mice, Inbred C57BL, Karyopherins metabolism, RNA, Messenger metabolism, RNA, Messenger genetics
- Abstract
Chronic obstructive pulmonary disease (COPD) poses a significant health threat characterized by lung inflammation primarily triggered by pulmonary monocytes. Despite the centrality of inflammation in COPD, the regulatory mechanisms governing this response remain elusive, presenting a challenge for anti-inflammatory interventions. In this study, we assessed the expression of exportins in COPD mouse models, revealing a notable upregulation of XPO6 in the mouse lung (P = 0.0011). Intriguingly, we observed a consistent upregulation of XPO6 in pulmonary monocytes from both human and mouse COPD subjects (P < 0.0001). Furthermore, in human lung tissue, XPO6 expression exhibited a positive correlation with TLR2 expression (P = 0). In vitro investigations demonstrated that XPO6 enhances TLR2 expression, activating the MyD88/NF-κB inflammatory signaling pathway. This activation, in turn, promotes the secretion of pro-inflammatory cytokines such as TNFα, IL-6, and IL-1β in monocytes. Mechanistically, XPO6 facilitates the nuclear export of TLR2 mRNA, ensuring its stability and subsequent protein expression in monocytes. In conclusion, our findings unveil that the upregulation of XPO6 in COPD pulmonary monocytes activates the MyD88/NF-κB inflammatory signaling pathway by facilitating the nuclear export of TLR2 mRNA, thereby identifying XPO6 as a promising therapeutic target for anti-inflammatory interventions in COPD., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)
- Published
- 2024
- Full Text
- View/download PDF
36. circLIFR-007 reduces liver metastasis via promoting hnRNPA1 nuclear export and YAP phosphorylation in breast cancer.
- Author
-
Zhang Y, Tan Y, Yuan J, Tang H, Zhang H, Tang Y, Xie Y, Wu L, Xie J, Xiao X, Li Y, and Kong Y
- Subjects
- Humans, Female, Phosphorylation, Animals, Mice, Cell Line, Tumor, Gene Expression Regulation, Neoplastic, Adaptor Proteins, Signal Transducing metabolism, Adaptor Proteins, Signal Transducing genetics, Active Transport, Cell Nucleus, Mice, Nude, Cell Proliferation, Mice, Inbred BALB C, MCF-7 Cells, Breast Neoplasms pathology, Breast Neoplasms metabolism, Breast Neoplasms genetics, Liver Neoplasms secondary, Liver Neoplasms metabolism, Liver Neoplasms genetics, YAP-Signaling Proteins metabolism, Heterogeneous Nuclear Ribonucleoprotein A1 metabolism, Heterogeneous Nuclear Ribonucleoprotein A1 genetics, RNA, Circular genetics, RNA, Circular metabolism, Transcription Factors metabolism, Transcription Factors genetics
- Abstract
Cancer metastasis is the major cause of death in patients with breast cancer (BC). The liver is a common site of breast cancer metastasis, and the 5-year survival rate of patients with breast cancer liver metastases (BCLMs) is only about 8.5 %. CircRNAs are involved in a variety of cancer-related pathological behaviors, and their unique structure and resistance to RNA degradation enable them to serve as ideal diagnostic biomarkers and therapeutic targets. Therefore, it is important to investigate the role and molecular mechanism of circRNAs in cancer metastasis. CircLIFR-007 was identified as a critical circular RNA in BC metastasis by circRNAs microarray and qRT-PCR experiment. Cell function assays were performed to explore the effect of circLIFR-007 in breast cancer cells. Experiments in vivo validated the function of circLIFR-007. Several molecular assays were performed to investigate the underlying mechanisms. We found that circLIFR-007 acted as a negative controller in breast cancer liver metastasis. CircLIFR-007 upregulates the phosphorylation level of YAP by exporting hnRNPA1 to promote the combination between hnRNPA1 and YAP in the cytoplasm. Overexpression of circLIFR-007 suppressed the expression of liver metastasis-related proteins, SREBF1 and SNAI1, which were regulated by transcription factor YAP. Functionally, circLIFR-007 inhibits the proliferation and metastasis of breast cancer cells both in vivo and in vitro., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. None., (Copyright © 2024. Published by Elsevier B.V.)
- Published
- 2024
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37. Wg/Wnt-signaling-induced nuclear translocation of β-catenin is attenuated by a β-catenin peptide through its interference with the IFT-A complex.
- Author
-
Vuong LT and Mlodzik M
- Subjects
- Animals, Humans, Wnt1 Protein metabolism, Wnt1 Protein genetics, Active Transport, Cell Nucleus, Drosophila melanogaster metabolism, Peptides metabolism, Peptides pharmacology, Protein Binding, Amino Acid Sequence, Transcription Factors, beta Catenin metabolism, Drosophila Proteins metabolism, Wnt Signaling Pathway, Cell Nucleus metabolism, Armadillo Domain Proteins metabolism, Armadillo Domain Proteins genetics
- Abstract
Wnt/Wingless (Wg) signaling is critical in development and disease, including cancer. Canonical Wnt signaling is mediated by β-catenin/Armadillo (Arm in Drosophila) transducing signals to the nucleus, with IFT-A/Kinesin 2 complexes promoting nuclear translocation of β-catenin/Arm. Here, we demonstrate that a conserved small N-terminal Arm
34 - 87 /β-catenin peptide binds to IFT140, acting as a dominant interference tool to attenuate Wg/Wnt signaling in vivo. Arm34 - 87 expression antagonizes endogenous Wnt/Wg signaling, resulting in the reduction of its target expression. Arm34 - 87 inhibits Wg/Wnt signaling by interfering with nuclear translocation of endogenous Arm/β-catenin, and this can be modulated by levels of wild-type β-catenin or IFT140, with the Arm34 - 87 effect being enhanced or suppressed. Importantly, this mechanism is conserved in mammals with the equivalent β-catenin24 - 79 peptide blocking nuclear translocation and pathway activation, including in cancer cells. Our work indicates that Wnt signaling can be regulated by a defined N-terminal β-catenin peptide and thus might serve as an entry point for therapeutic applications to attenuate Wnt/β-catenin signaling., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)- Published
- 2024
- Full Text
- View/download PDF
38. Endosome mediated nucleocytoplasmic trafficking and endomembrane allocation is crucial to polyglutamine toxicity.
- Author
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Nan Y, Chen W, Chen F, Wei L, Zeng A, Lin X, Zhou W, Yang Y, and Li Q
- Subjects
- Animals, Cell Nucleus metabolism, Drosophila Proteins metabolism, Drosophila Proteins genetics, Active Transport, Cell Nucleus, Drosophila melanogaster metabolism, Drosophila melanogaster genetics, Machado-Joseph Disease metabolism, Machado-Joseph Disease genetics, Machado-Joseph Disease pathology, Enterocytes metabolism, Disease Models, Animal, Ataxin-3 metabolism, Ataxin-3 genetics, Drosophila metabolism, Peptides metabolism, Endosomes metabolism, Autophagy
- Abstract
Aggregation of aberrant proteins is a common pathological hallmark in neurodegeneration such as polyglutamine (polyQ) and other repeat-expansion diseases. Here through overexpression of ataxin3 C-terminal polyQ expansion in Drosophila gut enterocytes, we generated an intestinal obstruction model of spinocerebellar ataxia type3 (SCA3) and reported a new role of nuclear-associated endosomes (NAEs)-the delivery of polyQ to the nucleoplasm. In this model, accompanied by the prominently increased RAB5-positive NAEs are abundant nucleoplasmic reticulum enriched with polyQ, abnormal nuclear envelope invagination, significantly reduced endoplasmic reticulum, indicating dysfunctional nucleocytoplasmic trafficking and impaired endomembrane organization. Consistently, Rab5 but not Rab7 RNAi further decreased polyQ-related NAEs, inhibited endomembrane disorganization, and alleviated disease model. Interestingly, autophagic proteins were enriched in polyQ-related NAEs and played non-canonical autophagic roles as genetic manipulation of autophagic molecules exhibited differential impacts on NAEs and SCA3 toxicity. Namely, the down-regulation of Atg1 or Atg12 mitigated while Atg5 RNAi aggravated the disease phenotypes both in Drosophila intestines and compound eyes. Our findings, therefore, provide new mechanistic insights and underscore the fundamental roles of endosome-centered nucleocytoplasmic trafficking and homeostatic endomembrane allocation in the pathogenesis of polyQ diseases., (© 2024. The Author(s).)
- Published
- 2024
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39. Altered nucleocytoplasmic export of adenosine-rich circRNAs by PABPC1 contributes to neuronal function.
- Author
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Cao SM, Wu H, Yuan GH, Pan YH, Zhang J, Liu YX, Li S, Xu YF, Wei MY, Yang L, and Chen LL
- Subjects
- Humans, Animals, Cell Line, Cell Differentiation, Cytoplasm metabolism, Prosencephalon metabolism, RNA, Circular metabolism, RNA, Circular genetics, Neurons metabolism, Active Transport, Cell Nucleus, Adenosine metabolism, Cell Nucleus metabolism, Neurogenesis, Poly(A)-Binding Protein I metabolism, Poly(A)-Binding Protein I genetics, RNA metabolism, RNA genetics
- Abstract
Circular RNAs (circRNAs) are upregulated during neurogenesis. Where and how circRNAs are localized and what roles they play during this process have remained elusive. Comparing the nuclear and cytoplasmic circRNAs between H9 cells and H9-derived forebrain (FB) neurons, we identify that a subset of adenosine (A)-rich circRNAs are restricted in H9 nuclei but exported to cytosols upon differentiation. Such a subcellular relocation of circRNAs is modulated by the poly(A)-binding protein PABPC1. In the H9 nucleus, newly produced (A)-rich circRNAs are bound by PABPC1 and trapped by the nuclear basket protein TPR to prevent their export. Modulating (A)-rich motifs in circRNAs alters their subcellular localization, and introducing (A)-rich circRNAs in H9 cytosols results in mRNA translation suppression. Moreover, decreased nuclear PABPC1 upon neuronal differentiation enables the export of (A)-rich circRNAs, including circRTN4(2,3), which is required for neurite outgrowth. These findings uncover subcellular localization features of circRNAs, linking their processing and function during neurogenesis., Competing Interests: Declaration of interests L.-L.C. is a member of the Molecular Cell advisory board and a scientific co-founder of RiboX Therapeutics., (Copyright © 2024 Elsevier Inc. All rights reserved.)
- Published
- 2024
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40. Respiratory syncytial virus NS1 inhibits anti-viral Interferon-α-induced JAK/STAT signaling, by limiting the nuclear translocation of STAT1.
- Author
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Efstathiou C, Zhang Y, Kandwal S, Fayne D, Molloy EJ, and Stevenson NJ
- Subjects
- Humans, Janus Kinases metabolism, Cell Nucleus metabolism, Phosphorylation, Active Transport, Cell Nucleus, Cell Line, STAT1 Transcription Factor metabolism, Signal Transduction, Interferon-alpha metabolism, Interferon-alpha pharmacology, Interferon-alpha immunology, Respiratory Syncytial Virus, Human immunology, Respiratory Syncytial Virus, Human physiology, Viral Nonstructural Proteins metabolism, Respiratory Syncytial Virus Infections immunology, Respiratory Syncytial Virus Infections metabolism, Respiratory Syncytial Virus Infections virology
- Abstract
Human respiratory viruses are the most prevalent cause of disease in humans, with the highly infectious RSV being the leading cause of infant bronchiolitis and viral pneumonia. Responses to type I IFNs are the primary defense against viral infection. However, RSV proteins have been shown to antagonize type I IFN-mediated antiviral innate immunity, specifically dampening intracellular IFN signaling. Respiratory epithelial cells are the main target for RSV infection. In this study, we found RSV-NS1 interfered with the IFN-α JAK/STAT signaling pathway of epithelial cells. RSV-NS1 expression significantly enhanced IFN-α-mediated phosphorylation of STAT1, but not pSTAT2; and neither STAT1 nor STAT2 total protein levels were affected by RSV-NS1. However, expression of RSV-NS1 significantly reduced ISRE and GAS promoter activity and anti-viral IRG expression. Further mechanistic studies demonstrated RSV-NS1 bound STAT1, with protein modeling indicating a possible interaction site between STAT1 and RSV-NS1. Nuclear translocation of STAT1 was reduced in the presence of RSV-NS1. Additionally, STAT1's interaction with the nuclear transport adapter protein, KPNA1, was also reduced, suggesting a mechanism by which RSV blocks STAT1 nuclear translocation. Indeed, reducing STAT1's access to the nucleus may explain RSV's suppression of IFN JAK/STAT promoter activation and antiviral gene induction. Taken together these results describe a novel mechanism by which RSV controls antiviral IFN-α JAK/STAT responses, which enhances our understanding of RSV's respiratory disease progression., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Efstathiou, Zhang, Kandwal, Fayne, Molloy and Stevenson.)
- Published
- 2024
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41. Nucleoporin Nup358 drives the differentiation of myeloid-biased multipotent progenitors by modulating HDAC3 nuclear translocation.
- Author
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Guglielmi V, Lam D, and D'Angelo MA
- Subjects
- Animals, Mice, Molecular Chaperones metabolism, Molecular Chaperones genetics, Myeloid Progenitor Cells metabolism, Myeloid Progenitor Cells cytology, Active Transport, Cell Nucleus, Cell Nucleus metabolism, Multipotent Stem Cells metabolism, Multipotent Stem Cells cytology, Myeloid Cells metabolism, Myeloid Cells cytology, Sumoylation, Myelopoiesis genetics, Histone Deacetylases metabolism, Histone Deacetylases genetics, Nuclear Pore Complex Proteins metabolism, Nuclear Pore Complex Proteins genetics, Cell Differentiation, Mice, Knockout
- Abstract
Nucleoporins, the components of nuclear pore complexes (NPCs), can play cell type- and tissue-specific functions. Yet, the physiological roles and mechanisms of action for most NPC components have not yet been established. We report that Nup358, a nucleoporin linked to several myeloid disorders, is required for the developmental progression of early myeloid progenitors. We found that Nup358 ablation in mice results in the loss of myeloid-committed progenitors and mature myeloid cells and the accumulation of myeloid-primed multipotent progenitors (MPPs) in bone marrow. Accumulated MPPs in Nup358 knockout mice are greatly restricted to megakaryocyte/erythrocyte-biased MPP2, which fail to progress into committed myeloid progenitors. Mechanistically, we found that Nup358 is required for histone deacetylase 3 (HDAC3) nuclear import and function in MPP2 cells and established that this nucleoporin regulates HDAC3 nuclear translocation in a SUMOylation-independent manner. Our study identifies a critical function for Nup358 in myeloid-primed MPP2 differentiation and uncovers an unexpected role for NPCs in the early steps of myelopoiesis.
- Published
- 2024
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42. WW domains form a folded type of nuclear localization signal to guide YAP1 nuclear import.
- Author
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Yang Y, Wu M, Pan Y, Hua Y, He X, Li X, Wang J, and Gan X
- Subjects
- Humans, Active Transport, Cell Nucleus, Proteins metabolism, WW Domains, alpha Karyopherins metabolism, beta Karyopherins metabolism, Cell Nucleus metabolism, Nuclear Localization Signals genetics, Nuclear Localization Signals metabolism, YAP-Signaling Proteins chemistry, YAP-Signaling Proteins metabolism
- Abstract
The nuclear translocation of YAP1 is significantly implicated in the proliferation, stemness, and metastasis of cancer cells. Although the molecular basis underlying YAP1 subcellular distribution has been extensively explored, it remains to be elucidated how the nuclear localization signal guides YAP1 to pass through the nuclear pore complex. Here, we define a globular type of nuclear localization signal composed of folded WW domains, named as WW-NLS. It directs YAP1 nuclear import through the heterodimeric nuclear transport receptors KPNA-KPNB1, bypassing the canonical nuclear localization signal that has been well documented in KPNA/KPNB1-mediated nuclear import. Strikingly, competitive interference with the function of the WW-NLS significantly attenuates YAP1 nuclear translocation and damages stemness gene activation and sphere formation in malignant breast cancer cells. Our findings elucidate a novel globular type of nuclear localization signal to facilitate nuclear entry of WW-containing proteins including YAP1., (© 2024 Yang et al.)
- Published
- 2024
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43. Vaccinia virus subverts xenophagy through phosphorylation and nuclear targeting of p62.
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Krause M, Samolej J, Yakimovich A, Kriston-Vizi J, Huttunen M, Lara-Reyna S, Frickel EM, and Mercer J
- Subjects
- Humans, Active Transport, Cell Nucleus, HEK293 Cells, HeLa Cells, Nuclear Proteins metabolism, Nuclear Proteins genetics, Phosphorylation, Vaccinia metabolism, Vaccinia virology, Vaccinia genetics, Virus Replication, Autophagy, Cell Nucleus metabolism, Cell Nucleus virology, Sequestosome-1 Protein metabolism, Sequestosome-1 Protein genetics, Vaccinia virus metabolism, Vaccinia virus genetics
- Abstract
Autophagy is an essential degradation program required for cell homeostasis. Among its functions is the engulfment and destruction of cytosolic pathogens, termed xenophagy. Not surprisingly, many pathogens use various strategies to circumvent or co-opt autophagic degradation. For poxviruses, it is known that infection activates autophagy, which however is not required for successful replication. Even though these complex viruses replicate exclusively in the cytoplasm, autophagy-mediated control of poxvirus infection has not been extensively explored. Using the prototypic poxvirus, vaccinia virus (VACV), we show that overexpression of the xenophagy receptors p62, NDP52, and Tax1Bp1 restricts poxvirus infection. While NDP52 and Tax1Bp1 were degraded, p62 initially targeted cytoplasmic virions before being shunted to the nucleus. Nuclear translocation of p62 was dependent upon p62 NLS2 and correlated with VACV kinase mediated phosphorylation of p62 T269/S272. This suggests that VACV targets p62 during the early stages of infection to avoid destruction and further implies that poxviruses exhibit multi-layered control of autophagy to facilitate cytoplasmic replication., (© 2024 Krause et al.)
- Published
- 2024
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44. The mRNA-capping enzyme localizes to stress granules in the cytoplasm and maintains cap homeostasis of target mRNAs.
- Author
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Gayen A, Mukherjee A, Kumar K, Majumder S, Chakrabarti S, and Mukherjee C
- Subjects
- Humans, RNA Caps metabolism, Arsenites pharmacology, Oxidative Stress, Active Transport, Cell Nucleus, RNA Nucleotidyltransferases metabolism, RNA Nucleotidyltransferases genetics, Sodium Compounds pharmacology, Exportin 1 Protein, Karyopherins metabolism, Karyopherins genetics, Receptors, Cytoplasmic and Nuclear metabolism, Receptors, Cytoplasmic and Nuclear genetics, Cytoplasmic Granules metabolism, RNA Stability, Cell Nucleus metabolism, Cell Line, Tumor, Nucleotidyltransferases, RNA, Messenger metabolism, RNA, Messenger genetics, Stress Granules metabolism, Homeostasis, Cytoplasm metabolism
- Abstract
The model of RNA stability has undergone a transformative shift with the revelation of a cytoplasmic capping activity that means a subset of transcripts are recapped autonomously of their nuclear counterparts. The present study demonstrates nucleo-cytoplasmic shuttling of the mRNA-capping enzyme (CE, also known as RNA guanylyltransferase and 5'-phosphatase; RNGTT), traditionally acknowledged for its nuclear localization and functions, elucidating its contribution to cytoplasmic capping activities. A unique nuclear export sequence in CE mediates XPO1-dependent nuclear export of CE. Notably, during sodium arsenite-induced oxidative stress, cytoplasmic CE (cCE) congregates within stress granules (SGs). Through an integrated approach involving molecular docking and subsequent co-immunoprecipitation, we identify eIF3b, a constituent of SGs, as an interactive associate of CE, implying that it has a potential role in guiding cCE to SGs. We measured the cap status of specific mRNA transcripts from U2OS cells that were non-stressed, stressed and recovered from stress, which indicated that cCE-target transcripts lost their caps during stress but remarkably regained cap stability during the recovery phase. This comprehensive study thus uncovers a novel facet of cytoplasmic CE, which facilitates cellular recovery from stress by maintaining cap homeostasis of target mRNAs., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2024. Published by The Company of Biologists Ltd.)
- Published
- 2024
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- View/download PDF
45. Asymmetric post-translational modifications regulate the nuclear translocation of STAT3 homodimers in response to leukemia inhibitory factor.
- Author
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Diallo M, Pimenta C, Murtinheira F, Martins-Alves D, Pinto FR, da Costa AA, Letra-Vilela R, Martin V, Rodriguez C, Rodrigues MS, and Herrera F
- Subjects
- Humans, Active Transport, Cell Nucleus, HeLa Cells, Phosphorylation, Protein Transport drug effects, Cell Nucleus metabolism, Leukemia Inhibitory Factor metabolism, Protein Multimerization, Protein Processing, Post-Translational, STAT3 Transcription Factor metabolism
- Abstract
STAT3 is a pleiotropic transcription factor overactivated in 70% of solid tumours. We have recently reported that inactivating mutations on residues susceptible to post-translational modifications (PTMs) in only one of the monomers (i.e. asymmetric) caused changes in the cellular distribution of STAT3 homodimers. Here, we used more controlled experimental conditions, i.e. without the interference of endogenous STAT3 (STAT3-/- HeLa cells) and in the presence of a defined cytokine stimulus (Leukemia Inhibitory Factor, LIF), to provide further evidence that asymmetric PTMs affect the nuclear translocation of STAT3 homodimers. Time-lapse microscopy for 20 min after LIF stimulation showed that S727 dephosphorylation (S727A) and K685 inactivation (K685R) slightly enhanced the nuclear translocation of STAT3 homodimers, while K49 inactivation (K49R) delayed STAT3 nuclear translocation. Our findings suggest that asymmetrically modified STAT3 homodimers could be a new level of STAT3 regulation and, therefore, a potential target for cancer therapy., (© 2023. The Author(s).)
- Published
- 2024
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46. Mammalian reovirus µ1 protein attenuates RIG-I and MDA5-mediated signaling transduction by blocking IRF3 phosphorylation and nuclear translocation.
- Author
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Wu B, Li D, Bai H, Mo R, Li H, Xie J, Zhang X, Yang Y, Li H, Idris A, Li X, and Feng R
- Subjects
- Animals, Humans, Active Transport, Cell Nucleus, Cell Nucleus metabolism, HEK293 Cells, Immunity, Innate immunology, Interferon-beta metabolism, Interferon-beta immunology, Phosphorylation, Protein Serine-Threonine Kinases, Reoviridae Infections immunology, Viral Proteins metabolism, DEAD Box Protein 58 metabolism, Interferon Regulatory Factor-3 metabolism, Interferon-Induced Helicase, IFIH1 metabolism, Interferon-Induced Helicase, IFIH1 genetics, Orthoreovirus, Mammalian immunology, Orthoreovirus, Mammalian physiology, Receptors, Immunologic, Signal Transduction immunology, Capsid Proteins metabolism
- Abstract
Mammalian reovirus (MRV) is a non-enveloped, gene segmented double-stranded RNA (dsRNA) virus. It is an important zoonotic pathogen that infects many mammals and vertebrates that act as natural hosts and causes respiratory and digestive tract diseases. Studies have reported that RIG-I and MDA5 in the innate immune cytoplasmic RNA-sensing RIG-like receptor (RLR) signaling pathway can recognize dsRNA from MRV and promote antiviral type I interferon (IFN) responses. However, the mechanism by which many MRV-encoded proteins evade the host innate immune response remains unclear. Here, we show that exogenous μ1 protein promoted the proliferation of MRV in vitro, while knockdown of MRV μ1 protein expression by shRNA could impair MRV proliferation. Specifically, μ1 protein inhibited MRV or poly(I:C)-induced IFN-β expression, and attenuated RIG-I/MDA5-mediated signaling axis transduction during MRV infection. Importantly, we found that μ1 protein significantly decreased IFN-β mRNA expression induced by MDA5, RIG-I, MAVS, TBK1, IRF3(5D), and degraded the protein expression of exogenous MDA5, RIG-I, MAVS, TBK1 and IRF3 via the proteasomal and lysosomal pathways. Additionally, we show that μ1 protein can physically interact with MDA5, RIG-I, MAVS, TBK1, and IRF3 and attenuate the RIG-I/MDA5-mediated signaling cascades by blocking the phosphorylation and nuclear translocation of IRF3. In conclusion, our findings reveal that MRV outer capsid protein μ1 is a key factor in antagonizing RLRs signaling cascades and provide new strategies for effective prevention and treatment of MRV infection., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)
- Published
- 2024
- Full Text
- View/download PDF
47. Global control of RNA polymerase II.
- Author
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Gillis A and Berry S
- Subjects
- Humans, Gene Expression Regulation, Cell Nucleus metabolism, RNA, Messenger metabolism, RNA, Messenger genetics, Active Transport, Cell Nucleus, Animals, RNA Polymerase II metabolism, Transcription, Genetic
- Abstract
RNA polymerase II (Pol II) is the multi-protein complex responsible for transcribing all protein-coding messenger RNA (mRNA). Most research on gene regulation is focused on the mechanisms controlling which genes are transcribed when, or on the mechanics of transcription. How global Pol II activity is determined receives comparatively less attention. Here, we follow the life of a Pol II molecule from 'assembly of the complex' to nuclear import, enzymatic activity, and degradation. We focus on how Pol II spends its time in the nucleus, and on the two-way relationship between Pol II abundance and activity in the context of homeostasis and global transcriptional changes., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Scott Berry reports financial support was provided by Australian Research Council. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)
- Published
- 2024
- Full Text
- View/download PDF
48. Atomic force microscopy of spherical intermediates on the pathway to fibril formation of influenza A virus nuclear export protein.
- Author
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Koroleva ON, Kuzmina NV, Dubrovin EV, and Drutsa VL
- Subjects
- Active Transport, Cell Nucleus, Microscopy, Atomic Force, Amyloid metabolism, Nuclear Proteins, Influenza A virus metabolism
- Abstract
The nuclear export protein of the influenza A virus (NEP) is involved in many important processes of the virus life cycle. This makes it an attractive target for the treatment of a disease caused by a virus. Previously it has been shown, that recombinant variants of NEP are highly prone to aggregation in solution under various conditions with the formation of amyloid-like aggregates. In the present work, the amyloid nature of NEP aggregates was evidenced by Congo red binding assays. Atomic force microscopy has shown that NEP can form two types of spherical nanoparticles, which provide an alternative pathway for the formation of amyloid-like fibrils. Type I of these "fibrillogenic" spheres, formed under physiological conditions, represents the micelle-like particles with height 10-60 nm, which can generate worm-like flexible fibrils with the diameter 2.5-4.0 nm, length 20-500 nm and the Young's modulus ~73 MPa. Type II spherical aggregates with size of about 400-1000 nm, formed at elevated temperatures, includes fractions of drop-like and vesicle-like particles, generating more rigid amyloid-like fibrils with height of ~8 nm, and length of up to 2 μm. The hypothetical mechanism of fibril formation via nanospherical structures was suggested. RESEARCH HIGHLIGHTS: AFM has revealed two types of the influenza A virus nuclear export protein spherical aggregates. They provide an alternative pathway for the formation of amyloid-like fibrils. The mechanism of fibril formation via spherical structures is suggested., (© 2024 Wiley Periodicals LLC.)
- Published
- 2024
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- View/download PDF
49. Compound Danshen Dripping Pill effectively alleviates cGAS-STING-triggered diseases by disrupting STING-TBK1 interaction.
- Author
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Shi W, Xu G, Gao Y, Yang H, Liu T, Zhao J, Li H, Wei Z, Hou X, Chen Y, Wen J, Li C, Zhao J, Zhang P, Wang Z, Xiao X, and Bai Z
- Subjects
- Mice, Mice, Inbred C57BL, Immunity, Innate, Membrane Proteins agonists, Membrane Proteins metabolism, Signal Transduction, THP-1 Cells, Exodeoxyribonucleases genetics, Phosphoproteins genetics, Macrophages, Interferon-beta metabolism, Interferon Regulatory Factor-3 metabolism, Transcription Factor RelA metabolism, Active Transport, Cell Nucleus, Obesity drug therapy, Diet, High-Fat, Protein Serine-Threonine Kinases metabolism, Humans, Drugs, Chinese Herbal, Salvia miltiorrhiza chemistry, Camphanes, Panax notoginseng chemistry, Autoimmune Diseases drug therapy, Inflammation drug therapy
- Abstract
Background: The cyclic GMP-AMP synthase (cGAS)-stimulator of interferon (IFN) genes (STING) pathway is critical in the innate immune system and can be mobilized by cytosolic DNA. The various inflammatory and autoimmune diseases progression is highly correlated with aberrant cGAS-STING pathway activation. While some cGAS-STING pathway inhibitor were identified, there are no drugs that can be applied to the clinic. Compound Danshen Dripping Pill (CDDP) has been successfully used in clinic around the world, but the most common application is limited to cardiovascular disease. Therefore, the purpose of the present investigation was to examine whether CDDP inhibits the cGAS-STING pathway and could be used as a therapeutic agent for multiple cGAS-STING-triggered diseases., Methods: BMDMs, THP1 cells or Trex1
-/- BMDMs were stimulated with various cGAS-STING-agonists after pretreatment with CDDP to detect the function of CDDP on IFN-β and ISGs productionn. Next, we detect the influence on IRF3 and P65 nuclear translocation, STING oligomerization and STING-TBK1-IRF3 complex formation of CDDP. Additionally, the DMXAA-mediated activation mice model of cGAS-STING pathway was used to study the effects of CDDP. Trex1-/- mice model and HFD-mediated obesity model were established to clarify the efficacy of CDDP on inflammatory and autoimmune diseases., Results: CDDP efficacy suppressed the IRF3 phosphorylation or the generation of IFN-β, ISGs, IL-6 and TNF-α. Mechanistically, CDDP did not influence the STING oligomerization and IRF3-TBK1 and STING-IRF3 interaction, but remarkably eliminated the STING-TBK1 interaction, ultimately blocking the downstream responses. In addition, we also clarified that CDDP could suppress cGAS-STING pathway activation triggered by DMXAA, in vivo. Consistently, CDDP could alleviate multi-organ inflammatory responses in Trex1-/- mice model and attenuate the inflammatory disorders, incleding obesity-induced insulin resistance., Conclusion: CDDP is a specifically cGAS-STING pathway inhibitor. Furthermore, we provide novel mechanism for CDDP and discovered a clinical agent for the therapy of cGAS-STING-triggered inflammatory and autoimmune diseases., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier GmbH. All rights reserved.)- Published
- 2024
- Full Text
- View/download PDF
50. Gene alterations in the nuclear transport receptor superfamily: A study of head and neck cancer.
- Author
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Nguyen PT, Shimojukkoku Y, Kajiya Y, Oku Y, Tomishima A, Shima K, and Sasahira T
- Subjects
- Humans, Gene Expression Regulation, Neoplastic, Receptors, Cytoplasmic and Nuclear genetics, Receptors, Cytoplasmic and Nuclear metabolism, Squamous Cell Carcinoma of Head and Neck genetics, Squamous Cell Carcinoma of Head and Neck metabolism, Squamous Cell Carcinoma of Head and Neck pathology, Active Transport, Cell Nucleus, Cell Line, Tumor, Head and Neck Neoplasms genetics, Head and Neck Neoplasms metabolism, Head and Neck Neoplasms pathology
- Abstract
In cancer cells, the nuclear transport system is often disrupted, leading to abnormal localization of nuclear proteins and altered gene expression. This disruption can arise from various mechanisms such as mutations in genes that regulate nuclear transport, altered expression of transport proteins, and changes in nuclear envelope structure. Oncogenic protein build-up in the nucleus due to the disturbance in nuclear transport can also boost tumor growth and cell proliferation. In this study, we performed bioinformatic analyses of 23 key nuclear transport receptors using genomic and transcriptomic data from pancancer and head and neck squamous cell carcinoma (HNSCC) datasets from The Cancer Genome Atlas (TCGA) and Cancer Cell Line Encyclopedia and found that the total alteration frequency of 23 nuclear transport receptors in 2691 samples of the PCAWG Consortium was 42.1% and a high levels of genetic alterations was significantly associated with poor overall survival. Amplification was the most common type of genetic alterations, and results in the overexpression of nuclear transport receptors in HNSCC compared to normal tissues. Furthermore, our study revealed that seven out of eight cell cycle genes (CDK1, CDK2, CDK4, CDK6, CCNA1, CCNB1, and CCNE2) were significantly and positively correlated with nuclear transport receptor genes in TCGA pancancer and CCLE datasets. Additionally, functional enrichment analysis showed that nuclear transport receptor genes were mainly enriched in the adhesion junction, cell cycle, ERBB, MAPK, MTOR and WNT signaling pathways., Competing Interests: The authors have declared that no competing interest exist., (Copyright: © 2024 Nguyen et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)
- Published
- 2024
- Full Text
- View/download PDF
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