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1.
Sci Adv ; 10(23): eadn8963, 2024 Jun 07.
Article in English | MEDLINE | ID: mdl-38838144

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.


Subject(s)
Cell Differentiation , Histone Deacetylases , Mice, Knockout , Nuclear Pore Complex Proteins , Animals , Histone Deacetylases/metabolism , Histone Deacetylases/genetics , Nuclear Pore Complex Proteins/metabolism , Nuclear Pore Complex Proteins/genetics , 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
2.
Methods Mol Biol ; 2502: 69-80, 2022.
Article in English | MEDLINE | ID: mdl-35412231

ABSTRACT

In eukaryotic cells the nuclear envelope encloses the genome separating it from the rest of the cell. Nuclear pore complexes are large multi protein channels that perforate the nuclear envelope, connecting the nucleus and the cytoplasm. Besides controlling nucleocytoplasmic molecule exchange, nuclear pore complexes create a permeability barrier that defines the maximum size of molecules that can freely diffuse into the nucleus. Accumulating evidence indicate that the permeability barrier of the nucleus can vary in different cellular conditions, during aging and in disease. Here we provide a simple protocol to analyze changes in nuclear permeability in plasma membrane-permeabilized cells and isolated nuclei using fluorescent dextrans of different sizes and confocal microscopy. The methods described herein represent a valuable resource to researchers studying the function of nuclear pore complexes and the dynamics of nuclear permeability in different cell types and processes.


Subject(s)
Dextrans , Nuclear Pore , Animals , Cell Nucleus/metabolism , Dextrans/metabolism , Mammals , Nuclear Envelope/metabolism , Nuclear Pore/metabolism , Permeability
3.
Article in English | MEDLINE | ID: mdl-34312247

ABSTRACT

In eukaryotic cells, the genetic material is segregated inside the nucleus. This compartmentalization of the genome requires a transport system that allows cells to move molecules across the nuclear envelope, the membrane-based barrier that surrounds the chromosomes. Nuclear pore complexes (NPCs) are the central component of the nuclear transport machinery. These large protein channels penetrate the nuclear envelope, creating a passage between the nucleus and the cytoplasm through which nucleocytoplasmic molecule exchange occurs. NPCs are one of the largest protein assemblies of eukaryotic cells and, in addition to their critical function in nuclear transport, these structures also play key roles in many cellular processes in a transport-independent manner. Here we will review the current knowledge of the NPC structure, the cellular mechanisms that regulate their formation and maintenance, and we will provide a brief description of a variety of processes that NPCs regulate.


Subject(s)
Eukaryotic Cells , Nuclear Pore , Active Transport, Cell Nucleus/physiology , Eukaryota/genetics , Nuclear Envelope/metabolism , Nuclear Pore/genetics , Nuclear Pore/metabolism
4.
Life Sci Alliance ; 5(3)2022 03.
Article in English | MEDLINE | ID: mdl-34911810

ABSTRACT

Nuclear pore complexes, the channels connecting the nucleus with the cytoplasm, are built by multiple copies of ∼30 proteins called nucleoporins. Recent evidence has exposed that nucleoporins can play cell type-specific functions. Despite novel discoveries into the cellular functions of nucleoporins, their role in the regulation of mammalian tissue physiology remains mostly unexplored because of a limited number of nucleoporin mouse models. Here we show that ablation of Nup210/Gp210, a nucleoporin previously identified to play a role in myoblast differentiation and Zebrafish muscle maturation, is dispensable for skeletal muscle formation and growth in mice. We found that although primary satellite cells from Nup210 knockout mice can differentiate, these animals show delayed muscle repair after injury. Moreover, Nup210 knockout mice display an increased percentage of centrally nucleated fibers and abnormal fiber type distribution as they age. Muscle function experiments also exposed that Nup210 is required for muscle endurance during voluntary running. Our findings indicate that in mammals, Nup210 is important for the maintenance of skeletal muscle integrity and for proper muscle function providing novel insights into the in vivo roles of nuclear pore complex components.


Subject(s)
Muscles/metabolism , Nuclear Pore Complex Proteins/deficiency , Phenotype , Regeneration/genetics , Age Factors , Animals , Cell Differentiation , Fluorescent Antibody Technique , Gene Expression Regulation , Mice , Mice, Knockout , Muscle Development/genetics , Muscles/pathology , Satellite Cells, Skeletal Muscle/cytology , Satellite Cells, Skeletal Muscle/metabolism
5.
Cancer Discov ; 11(1): 176-193, 2021 01.
Article in English | MEDLINE | ID: mdl-32988961

ABSTRACT

Nuclear pore complexes (NPC) are the central mediators of nucleocytoplasmic transport. Increasing evidence shows that many cancer cells have increased numbers of NPCs and become addicted to the nuclear transport machinery. How reducing NPC numbers affects the physiology of normal and cancer cells and whether it could be exploited for cancer therapies has not been investigated. We report that inhibition of NPC formation, a process mostly restricted to proliferating cells, causes selective cancer cell death, prevents tumor growth, and induces tumor regression. Although cancer cells die in response to NPC assembly inhibition, normal cells undergo a reversible cell-cycle arrest that allows them to survive. Mechanistically, reducing NPC numbers results in multiple alterations contributing to cancer cell death, including abnormalities in nuclear transport, catastrophic alterations in gene expression, and the selective accumulation of DNA damage. Our findings uncover the NPC formation process as a novel targetable pathway in cancer cells. SIGNIFICANCE: Reducing NPC numbers in cancer cells induces death, prevents tumor growth, and results in tumor regression. Conversely, normal cells undergo a reversible cell-cycle arrest in response to inhibition of NPC assembly. These findings expose the potential of targeting NPC formation in cancer.This article is highlighted in the In This Issue feature, p. 1.


Subject(s)
Neoplasms , Nuclear Pore , Active Transport, Cell Nucleus , Cell Death , Humans , Neoplasms/drug therapy , Neoplasms/genetics , Neoplasms/metabolism , Nuclear Pore/metabolism , Nuclear Pore Complex Proteins/metabolism
6.
Development ; 147(23)2020 12 15.
Article in English | MEDLINE | ID: mdl-33323374

ABSTRACT

Nuclear pore complexes are multiprotein channels that span the nuclear envelope, which connects the nucleus to the cytoplasm. In addition to their main role in the regulation of nucleocytoplasmic molecule exchange, it has become evident that nuclear pore complexes and their components also have multiple transport-independent functions. In recent years, an increasing number of studies have reported the involvement of nuclear pore complex components in embryogenesis, cell differentiation and tissue-specific processes. Here, we review the findings that highlight the dynamic nature of nuclear pore complexes and their roles in many cell type-specific functions during development and tissue homeostasis.


Subject(s)
Cell Nucleus/genetics , Homeostasis/genetics , Nuclear Pore Complex Proteins/genetics , Nuclear Pore/genetics , Animals , Cell Differentiation/genetics , Cytoplasm/genetics , Embryonic Development/genetics , Humans , Nuclear Envelope/genetics , Organ Specificity/genetics
7.
Neuron ; 102(3): 509-511, 2019 05 08.
Article in English | MEDLINE | ID: mdl-31071281

ABSTRACT

Increasing evidence points to nuclear pore complexes as important regulators of cell fate and tissue homeostasis. A recent report by Liu et al. (2019) in this issue of Neuron uncovers that nucleoporin Seh1 is required for the expression of genes critical for oligodendrocyte differentiation and myelination.


Subject(s)
Nuclear Pore Complex Proteins , Nuclear Pore , Cell Differentiation , Oligodendroglia
8.
Nat Immunol ; 19(6): 594-605, 2018 06.
Article in English | MEDLINE | ID: mdl-29736031

ABSTRACT

Nuclear pore complexes (NPCs) are channels connecting the nucleus with the cytoplasm. We report that loss of the tissue-specific NPC component Nup210 causes a severe deficit of naïve CD4+ T cells. Nup210-deficient CD4+ T lymphocytes develop normally but fail to survive in the periphery. The decreased survival results from both an impaired ability to transmit tonic T cell receptor (TCR) signals and increased levels of Fas, which sensitize Nup210-/- naïve CD4+ T cells to Fas-mediated cell death. Mechanistically, Nup210 regulates these processes by modulating the expression of Cav2 (encoding Caveolin-2) and Jun at the nuclear periphery. Whereas the TCR-dependent and CD4+ T cell-specific upregulation of Cav2 is critical for proximal TCR signaling, cJun expression is required for STAT3-dependent repression of Fas. Our results uncover an unexpected role for Nup210 as a cell-intrinsic regulator of TCR signaling and T cell homeostasis and expose NPCs as key players in the adaptive immune system.


Subject(s)
CD4-Positive T-Lymphocytes/immunology , Homeostasis/immunology , Nuclear Pore Complex Proteins/immunology , Receptors, Antigen, T-Cell/immunology , Signal Transduction/immunology , Animals , CD4-Positive T-Lymphocytes/metabolism , Mice , Mice, Inbred C57BL , Mice, Transgenic , Nuclear Pore/immunology , Nuclear Pore/metabolism , Nuclear Pore Complex Proteins/metabolism , Receptors, Antigen, T-Cell/metabolism
10.
Nucleus ; 9(1): 142-148, 2018 01 01.
Article in English | MEDLINE | ID: mdl-29095096

ABSTRACT

Nuclear pore complexes (NPCs), the channels connecting the nucleus with the cytoplasm, are the largest protein structures of the nuclear envelope. In addition to their role in regulating nucleocytoplasmic transport, increasing evidence shows that these multiprotein structures play central roles in the regulation of gene activity. In light of recent discoveries, NPCs are emerging as scaffolds that mediate the regulation of specific gene sets at the nuclear periphery. The function of NPCs as genome organizers and hubs for transcriptional regulation provides additional evidence that the compartmentalization of genes and transcriptional regulators within the nuclear space is an important mechanism of gene expression regulation.


Subject(s)
Gene Expression Regulation , Nuclear Pore Complex Proteins/metabolism , Animals , Humans
11.
Dev Cell ; 41(5): 540-554.e7, 2017 06 05.
Article in English | MEDLINE | ID: mdl-28586646

ABSTRACT

Nuclear pore complexes (NPCs) are multiprotein channels connecting the nucleus with the cytoplasm. NPCs have been shown to have tissue-specific composition, suggesting that their function can be specialized. However, the physiological roles of NPC composition changes and their impacts on cellular processes remain unclear. Here we show that the addition of the Nup210 nucleoporin to NPCs during myoblast differentiation results in assembly of an Mef2C transcriptional complex required for efficient expression of muscle structural genes and microRNAs. We show that this NPC-localized complex is essential for muscle growth, myofiber maturation, and muscle cell survival and that alterations in its activity result in muscle degeneration. Our findings suggest that NPCs regulate the activity of functional gene groups by acting as scaffolds that promote the local assembly of tissue-specific transcription complexes and show how nuclear pore composition changes can be exploited to regulate gene expression at the nuclear periphery.


Subject(s)
Embryo, Nonmammalian/cytology , MEF2 Transcription Factors/metabolism , Muscle Development/genetics , Nuclear Pore Complex Proteins/metabolism , Nuclear Pore/physiology , Zebrafish Proteins/metabolism , Zebrafish/metabolism , Animals , Cell Differentiation , Cell Nucleus/genetics , Embryo, Nonmammalian/metabolism , MEF2 Transcription Factors/genetics , Nuclear Envelope/genetics , Nuclear Pore Complex Proteins/genetics , Zebrafish/growth & development , Zebrafish Proteins/genetics
12.
Semin Cell Dev Biol ; 68: 72-84, 2017 08.
Article in English | MEDLINE | ID: mdl-28506892

ABSTRACT

The study of the Nuclear Pore Complex (NPC), the proteins that compose it (nucleoporins), and the nucleocytoplasmic transport that it controls have revealed an unexpected layer to pathogenic disease onset and progression. Recent advances in the study of the regulation of NPC composition and function suggest that the precise control of this structure is necessary to prevent diseases from arising or progressing. Here we discuss the role of nucleoporins in a diverse set of diseases, many of which directly or indirectly increase in occurrence and severity as we age, and often shorten the human lifespan. NPC biology has been shown to play a direct role in these diseases and therefore in the process of healthy aging.


Subject(s)
Nuclear Pore Complex Proteins/metabolism , Nuclear Pore/metabolism , Aging , Disease , Humans
13.
Curr Opin Cell Biol ; 46: 26-32, 2017 06.
Article in English | MEDLINE | ID: mdl-28088069

ABSTRACT

Nuclear pore complexes (NPCs), are large multiprotein channels that penetrate the nuclear envelope connecting the nucleus to the cytoplasm. Accumulating evidence shows that besides their main role in regulating the exchange of molecules between these two compartments, NPCs and their components also play important transport-independent roles, including gene expression regulation, chromatin organization, DNA repair, RNA processing and quality control, and cell cycle control. Here, we will describe the recent findings about the role of these structures in the regulation of gene expression.


Subject(s)
Gene Expression Regulation , Nuclear Pore Complex Proteins/metabolism , Animals , Cell Nucleus/metabolism , Genome , Humans , Nuclear Envelope/metabolism , Nuclear Pore/metabolism , Yeasts/cytology , Yeasts/genetics
15.
Curr Opin Cell Biol ; 25(3): 378-86, 2013 Jun.
Article in English | MEDLINE | ID: mdl-23567027

ABSTRACT

Maintaining genome integrity is crucial for successful organismal propagation and for cell and tissue homeostasis. Several processes contribute to safeguarding the genomic information of cells. These include accurate replication of genetic information, detection and repair of DNA damage, efficient segregation of chromosomes, protection of chromosome ends, and proper organization of genome architecture. Interestingly, recent evidence shows that nuclear pore complexes, the channels connecting the nucleus with the cytoplasm, play important roles in these processes suggesting that these multiprotein platforms are key regulators of genome integrity.


Subject(s)
Cell Nucleus/metabolism , Genomic Instability , Nuclear Pore/metabolism , Animals , Cell Nucleus/genetics , Chromosomes/metabolism , Cytoplasm/genetics , DNA Damage , DNA Replication , Humans
16.
Nat Rev Mol Cell Biol ; 13(11): 687-99, 2012 Nov.
Article in English | MEDLINE | ID: mdl-23090414

ABSTRACT

Nuclear pore complexes (NPCs) are multiprotein aqueous channels that penetrate the nuclear envelope connecting the nucleus and the cytoplasm. NPCs consist of multiple copies of roughly 30 different proteins known as nucleoporins (NUPs). Due to their essential role in controlling nucleocytoplasmic transport, NPCs have traditionally been considered as structures of ubiquitous composition. The overall structure of the NPC is indeed conserved in all cells, but new evidence suggests that the protein composition of NPCs varies among cell types and tissues. Moreover, mutations in various nucleoporins result in tissue-specific diseases. These findings point towards a heterogeneity in NPC composition and function. This unexpected heterogeneity suggests that cells use a combination of different nucleoporins to assemble NPCs with distinct properties and specialized functions.


Subject(s)
Active Transport, Cell Nucleus , Nuclear Pore Complex Proteins/metabolism , Nuclear Pore/metabolism , Active Transport, Cell Nucleus/genetics , Animals , Cell Nucleus/metabolism , Cytoplasm/metabolism , Drosophila melanogaster/metabolism , Genetic Variation , Humans , Nuclear Envelope/metabolism , Nuclear Pore/chemistry , Nuclear Pore Complex Proteins/chemistry , Nuclear Pore Complex Proteins/genetics
17.
Dev Cell ; 22(2): 446-58, 2012 Feb 14.
Article in English | MEDLINE | ID: mdl-22264802

ABSTRACT

Nuclear pore complexes (NPCs) are built from ∼30 different proteins called nucleoporins or Nups. Previous studies have shown that several Nups exhibit cell-type-specific expression and that mutations in NPC components result in tissue-specific diseases. Here we show that a specific change in NPC composition is required for both myogenic and neuronal differentiation. The transmembrane nucleoporin Nup210 is absent in proliferating myoblasts and embryonic stem cells (ESCs) but becomes expressed and incorporated into NPCs during cell differentiation. Preventing Nup210 production by RNAi blocks myogenesis and the differentiation of ESCs into neuroprogenitors. We found that the addition of Nup210 to NPCs does not affect nuclear transport but is required for the induction of genes that are essential for cell differentiation. Our results identify a single change in NPC composition as an essential step in cell differentiation and establish a role for Nup210 in gene expression regulation and cell fate determination.


Subject(s)
Cell Differentiation , Embryonic Stem Cells/metabolism , Muscle Development/physiology , Neurons/metabolism , Nuclear Pore Complex Proteins/metabolism , Nuclear Pore/physiology , Active Transport, Cell Nucleus , Animals , Biomarkers/metabolism , Blotting, Western , Cell Proliferation , Embryonic Stem Cells/cytology , Gene Expression Profiling , Mice , Muscle Fibers, Skeletal/cytology , Muscle Fibers, Skeletal/metabolism , Myoblasts/cytology , Myoblasts/metabolism , Neurons/cytology , Nuclear Pore Complex Proteins/antagonists & inhibitors , Nuclear Pore Complex Proteins/genetics , Oligonucleotide Array Sequence Analysis , RNA, Messenger/genetics , RNA, Small Interfering/genetics , Real-Time Polymerase Chain Reaction
18.
Cell ; 136(2): 284-95, 2009 Jan 23.
Article in English | MEDLINE | ID: mdl-19167330

ABSTRACT

In dividing cells, nuclear pore complexes (NPCs) disassemble during mitosis and reassemble into the newly forming nuclei. However, the fate of nuclear pores in postmitotic cells is unknown. Here, we show that NPCs, unlike other nuclear structures, do not turn over in differentiated cells. While a subset of NPC components, like Nup153 and Nup50, are continuously exchanged, scaffold nucleoporins, like the Nup107/160 complex, are extremely long-lived and remain incorporated in the nuclear membrane during the entire cellular life span. Besides the lack of nucleoporin expression and NPC turnover, we discovered an age-related deterioration of NPCs, leading to an increase in nuclear permeability and the leaking of cytoplasmic proteins into the nucleus. Our finding that nuclear "leakiness" is dramatically accelerated during aging and that a subset of nucleoporins is oxidatively damaged in old cells suggests that the accumulation of damage at the NPC might be a crucial aging event.


Subject(s)
Cell Nucleus/physiology , Mitosis , Nuclear Pore/physiology , Animals , Caenorhabditis elegans , Down-Regulation , Mice , Nuclear Pore Complex Proteins/physiology , Rats
19.
Trends Cell Biol ; 18(10): 456-66, 2008 Oct.
Article in English | MEDLINE | ID: mdl-18786826

ABSTRACT

Nuclear pore complexes are large aqueous channels that penetrate the nuclear envelope, thereby connecting the nuclear interior with the cytoplasm. Until recently, these macromolecular complexes were viewed as static structures, the only function of which was to control the molecular trafficking between the two compartments. It has now become evident that this simplistic scenario is inaccurate and that nuclear pore complexes are highly dynamic multiprotein assemblies involved in diverse cellular processes ranging from the organization of the cytoskeleton to gene expression. In this review, we discuss the most recent developments in the nuclear-pore-complex field, focusing on the assembly, disassembly, maintenance and function of this macromolecular structure.


Subject(s)
Cell Nucleus/physiology , Nuclear Pore Complex Proteins/physiology , Nuclear Pore/physiology , Animals , Chromatin/physiology , Cytoskeleton/physiology , Humans , Mitosis/physiology , Nuclear Envelope/physiology , Nuclear Pore/chemistry , Nuclear Pore Complex Proteins/chemistry , Saccharomyces cerevisiae/metabolism
20.
J Virol ; 80(19): 9608-18, 2006 Oct.
Article in English | MEDLINE | ID: mdl-16973564

ABSTRACT

Adenoviruses are nonenveloped viruses with an approximately 36-kb double-stranded DNA genome that replicate in the nucleus. Protein VII, an abundant structural component of the adenovirus core that is strongly associated with adenovirus DNA, is imported into the nucleus contemporaneously with the adenovirus genome shortly after virus infection and may promote DNA import. In this study, we evaluated whether protein VII uses specific receptor-mediated mechanisms for import into the nucleus. We found that it contains potent nuclear localization signal (NLS) activity by transfection of cultured cells with protein VII fusion constructs and by microinjection of cells with recombinant protein VII fusions. We identified three NLS-containing regions in protein VII by deletion mapping and determined important NLS residues by site-specific mutagenesis. We found that recombinant protein VII and its NLS-containing domains strongly and specifically bind to importin alpha, importin beta, importin 7, and transportin, which are among the most abundant cellular nuclear import receptors. Moreover, these receptors can mediate the nuclear import of protein VII fusions in vitro in permeabilized cells. Considered together, these data support the hypothesis that protein VII is a major NLS-containing adaptor for receptor-mediated import of adenovirus DNA and that multiple import pathways are utilized to promote efficient nuclear entry of the viral genome.


Subject(s)
Adenoviridae/metabolism , Viral Core Proteins/metabolism , Active Transport, Cell Nucleus , Adenoviridae/chemistry , Adenoviridae/genetics , Animals , Cell Line , Chlorocebus aethiops , Cytosol/metabolism , Humans , Molecular Sequence Data , Mutation/genetics , Nuclear Localization Signals , Protein Binding , Viral Core Proteins/chemistry , Viral Core Proteins/genetics
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