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  1. Article ; Online: F-actin-rich territories coordinate apoptosome assembly and caspase activation during DNA damage-induced intrinsic apoptosis.

    King, Virginia L / Campellone, Kenneth G

    Molecular biology of the cell

    2023  Volume 34, Issue 5, Page(s) ar41

    Abstract: The actin cytoskeleton is a ubiquitous participant in cellular functions that maintain viability, but how it controls programmed cell death is not well understood. Here we show that in response to DNA damage, human cells form a juxtanuclear F-actin-rich ... ...

    Abstract The actin cytoskeleton is a ubiquitous participant in cellular functions that maintain viability, but how it controls programmed cell death is not well understood. Here we show that in response to DNA damage, human cells form a juxtanuclear F-actin-rich territory that coordinates the organized progression of apoptosome assembly to caspase activation. This cytoskeletal compartment is created by the actin nucleation factors JMY, WHAMM, and the Arp2/3 complex, and it excludes proteins that inhibit JMY and WHAMM activity. Within the territory, mitochondria undergo outer membrane permeabilization and JMY localization overlaps with punctate structures containing the core apoptosome components cytochrome
    MeSH term(s) Humans ; Actins/metabolism ; Caspase 3 ; Apoptosomes/metabolism ; Apoptosis/physiology ; Caspases/metabolism ; Actin Cytoskeleton/metabolism ; DNA Damage ; Membrane Proteins/metabolism ; Microtubule-Associated Proteins
    Chemical Substances Actins ; Caspase 3 (EC 3.4.22.-) ; Apoptosomes ; Caspases (EC 3.4.22.-) ; WHAMM protein, human ; Membrane Proteins ; Microtubule-Associated Proteins
    Language English
    Publishing date 2023-03-15
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural
    ZDB-ID 1098979-1
    ISSN 1939-4586 ; 1059-1524
    ISSN (online) 1939-4586
    ISSN 1059-1524
    DOI 10.1091/mbc.E22-04-0119
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    Zs.A 2853: Show issues Location:
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  2. Article: Autophagosome turnover requires Arp2/3 complex-mediated maintenance of lysosomal integrity.

    Theodore, Corey J / Wagner, Lianna H / Campellone, Kenneth G

    bioRxiv : the preprint server for biology

    2024  

    Abstract: Autophagy is an intracellular degradation process that maintains homeostasis, responds to stress, and plays key roles in the prevention of aging and disease. Autophagosome biogenesis, vesicle rocketing, and autolysosome tubulation are controlled by ... ...

    Abstract Autophagy is an intracellular degradation process that maintains homeostasis, responds to stress, and plays key roles in the prevention of aging and disease. Autophagosome biogenesis, vesicle rocketing, and autolysosome tubulation are controlled by multiple actin nucleation factors, but the impact of actin assembly on completion of the autophagic pathway is not well understood. Here we studied autophagosome and lysosome remodeling in fibroblasts harboring an inducible knockout (iKO) of the Arp2/3 complex, an essential actin nucleator. Arp2/3 complex ablation resulted in increased basal levels of autophagy receptors and lipidated membrane proteins from the LC3 and GABARAP families. Under both steady-state and starvation conditions, Arp2/3 iKO cells accumulated abnormally high numbers of autolysosomes, suggesting a defect in autophagic flux. The inability of Arp2/3 complex-deficient cells to complete autolysosome degradation and turnover is explained by the presence of damaged, leaky lysosomes. In cells treated with an acute lysosomal membrane-damaging agent, the Arp2/3-activating protein WHAMM is recruited to lysosomes, where Arp2/3 complex-dependent actin assembly is crucial for restoring intact lysosomal structure. These results establish the Arp2/3 complex as a central player late in the canonical autophagy pathway and reveal a new role for the actin nucleation machinery in maintaining lysosomal integrity.
    Language English
    Publishing date 2024-03-13
    Publishing country United States
    Document type Preprint
    DOI 10.1101/2024.03.12.584718
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article ; Online: Branching out in different directions: Emerging cellular functions for the Arp2/3 complex and WASP-family actin nucleation factors.

    Campellone, Kenneth G / Lebek, Nadine M / King, Virginia L

    European journal of cell biology

    2023  Volume 102, Issue 2, Page(s) 151301

    Abstract: The actin cytoskeleton impacts practically every function of a eukaryotic cell. Historically, the best-characterized cytoskeletal activities are in cell morphogenesis, motility, and division. The structural and dynamic properties of the actin ... ...

    Abstract The actin cytoskeleton impacts practically every function of a eukaryotic cell. Historically, the best-characterized cytoskeletal activities are in cell morphogenesis, motility, and division. The structural and dynamic properties of the actin cytoskeleton are also crucial for establishing, maintaining, and changing the organization of membrane-bound organelles and other intracellular structures. Such activities are important in nearly all animal cells and tissues, although distinct anatomical regions and physiological systems rely on different regulatory factors. Recent work indicates that the Arp2/3 complex, a broadly expressed actin nucleator, drives actin assembly during several intracellular stress response pathways. These newly described Arp2/3-mediated cytoskeletal rearrangements are coordinated by members of the Wiskott-Aldrich Syndrome Protein (WASP) family of actin nucleation-promoting factors. Thus, the Arp2/3 complex and WASP-family proteins are emerging as crucial players in cytoplasmic and nuclear activities including autophagy, apoptosis, chromatin dynamics, and DNA repair. Characterizations of the functions of the actin assembly machinery in such stress response mechanisms are advancing our understanding of both normal and pathogenic processes, and hold great promise for providing insights into organismal development and interventions for disease.
    MeSH term(s) Animals ; Wiskott-Aldrich Syndrome Protein Family/metabolism ; Actins/metabolism ; Actin-Related Protein 2-3 Complex/metabolism ; Actin Cytoskeleton/metabolism ; Cytoskeleton/metabolism ; Wiskott-Aldrich Syndrome Protein/genetics ; Wiskott-Aldrich Syndrome Protein/metabolism ; Actin-Related Protein 3/metabolism
    Chemical Substances Wiskott-Aldrich Syndrome Protein Family ; Actins ; Actin-Related Protein 2-3 Complex ; Wiskott-Aldrich Syndrome Protein ; Actin-Related Protein 3
    Language English
    Publishing date 2023-03-02
    Publishing country Germany
    Document type Journal Article
    ZDB-ID 391967-5
    ISSN 1618-1298 ; 0070-2463 ; 0171-9335
    ISSN (online) 1618-1298
    ISSN 0070-2463 ; 0171-9335
    DOI 10.1016/j.ejcb.2023.151301
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    Zs.A 667: Show issues Location:
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  4. Article ; Online: WHAMM functions in kidney reabsorption and polymerizes actin to promote autophagosomal membrane closure and cargo sequestration.

    Coulter, Alyssa M / Cortés, Valerie / Theodore, Corey J / Cianciolo, Rachel E / Korstanje, Ron / Campellone, Kenneth G

    Molecular biology of the cell

    2024  , Page(s) mbcE24010025

    Abstract: The actin cytoskeleton is essential for many functions of eukaryotic cells, but the factors that nucleate actin assembly are not well understood at the organismal level or in the context of disease. To explore the function of the actin nucleation factor ... ...

    Abstract The actin cytoskeleton is essential for many functions of eukaryotic cells, but the factors that nucleate actin assembly are not well understood at the organismal level or in the context of disease. To explore the function of the actin nucleation factor WHAMM in mice, we examined how
    Language English
    Publishing date 2024-04-10
    Publishing country United States
    Document type Journal Article
    ZDB-ID 1098979-1
    ISSN 1939-4586 ; 1059-1524
    ISSN (online) 1939-4586
    ISSN 1059-1524
    DOI 10.1091/mbc.E24-01-0025
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  5. Article: WHAMM functions in kidney reabsorption and polymerizes actin to promote autophagosomal membrane closure and cargo sequestration.

    Coulter, Alyssa M / Cortés, Valerie / Theodore, Corey J / Cianciolo, Rachel E / Korstanje, Ron / Campellone, Kenneth G

    bioRxiv : the preprint server for biology

    2024  

    Abstract: The actin cytoskeleton is essential for many functions of eukaryotic cells, but the factors that nucleate actin assembly are not well understood at the organismal level or in the context of disease. To explore the function of the actin nucleation factor ... ...

    Abstract The actin cytoskeleton is essential for many functions of eukaryotic cells, but the factors that nucleate actin assembly are not well understood at the organismal level or in the context of disease. To explore the function of the actin nucleation factor WHAMM in mice, we examined how
    Language English
    Publishing date 2024-01-23
    Publishing country United States
    Document type Preprint
    DOI 10.1101/2024.01.22.576497
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  6. Article ; Online: Lambda Red-Mediated Recombination in Shiga Toxin-Producing Escherichia coli.

    Campellone, Kenneth G / Coulter, Alyssa M

    Methods in molecular biology (Clifton, N.J.)

    2021  Volume 2291, Page(s) 145–162

    Abstract: The bacteriophage Lambda (λ) "Red" recombination system has enabled the development of efficient methods for engineering bacterial chromosomes. This system has been particularly important to the field of bacterial pathogenesis, where it has advanced the ... ...

    Abstract The bacteriophage Lambda (λ) "Red" recombination system has enabled the development of efficient methods for engineering bacterial chromosomes. This system has been particularly important to the field of bacterial pathogenesis, where it has advanced the study of virulence factors from Shiga toxin-producing and enteropathogenic Escherichia coli (STEC and EPEC). Transient plasmid-driven expression of Lambda Red allows homologous recombination between PCR-derived linear DNA substrates and target loci in the STEC/EPEC chromosomes. Red-associated techniques can be used to create individual gene knockouts, generate deletions of large pathogenicity islands, and make markerless allelic exchanges. This chapter describes specific strategies and procedures for performing Lambda Red-mediated genome engineering in STEC.
    MeSH term(s) Bacteriophage lambda/genetics ; Bacteriophage lambda/metabolism ; Enteropathogenic Escherichia coli/genetics ; Enteropathogenic Escherichia coli/metabolism ; Escherichia coli Infections/genetics ; Escherichia coli Infections/metabolism ; Recombination, Genetic ; Shiga-Toxigenic Escherichia coli/genetics ; Shiga-Toxigenic Escherichia coli/metabolism ; Viral Proteins/genetics ; Viral Proteins/metabolism
    Chemical Substances Viral Proteins
    Language English
    Publishing date 2021-03-11
    Publishing country United States
    Document type Journal Article
    ISSN 1940-6029
    ISSN (online) 1940-6029
    DOI 10.1007/978-1-0716-1339-9_6
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  7. Article ; Online: Adding SNX to the mix: SNX9 drives filopodia biogenesis.

    Lebek, Nadine M / Campellone, Kenneth G

    The Journal of cell biology

    2020  Volume 219, Issue 4

    Abstract: Filopodia are actin-rich protrusions important for sensing and responding to the extracellular environment, but the repertoire of factors required for filopodia formation is only partially understood. Jarsch et al. (2020. J. Cell. Biol. https://doi.org/ ... ...

    Abstract Filopodia are actin-rich protrusions important for sensing and responding to the extracellular environment, but the repertoire of factors required for filopodia formation is only partially understood. Jarsch et al. (2020. J. Cell. Biol. https://doi.org/10.1083/jcb.201909178) combine an in vitro system of filopodia biogenesis with a phage display screen to show that SNX9 drives filopodial assembly.
    MeSH term(s) Actins ; Pseudopodia
    Chemical Substances Actins
    Language English
    Publishing date 2020-04-09
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Comment
    ZDB-ID 218154-x
    ISSN 1540-8140 ; 0021-9525
    ISSN (online) 1540-8140
    ISSN 0021-9525
    DOI 10.1083/jcb.202002086
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    Ub I Zs.190: Show issues Location:
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  8. Article ; Online: Branching out in different directions: Emerging cellular functions for the Arp2/3 complex and WASP-family actin nucleation factors

    Campellone, Kenneth G / Lebek, Nadine M / King, Virginia L

    European Journal of Cell Biology. 2023 June, v. 102, no. 2, p. 151301

    2023  , Page(s) 151301

    Abstract: The actin cytoskeleton impacts practically every function of a eukaryotic cell. Historically, the best-characterized cytoskeletal activities are in cell morphogenesis, motility, and division. The structural and dynamic properties of the actin ... ...

    Abstract The actin cytoskeleton impacts practically every function of a eukaryotic cell. Historically, the best-characterized cytoskeletal activities are in cell morphogenesis, motility, and division. The structural and dynamic properties of the actin cytoskeleton are also crucial for establishing, maintaining, and changing the organization of membrane-bound organelles and other intracellular structures. Such activities are important in nearly all animal cells and tissues, although distinct anatomical regions and physiological systems rely on different regulatory factors. Recent work indicates that the Arp2/3 complex, a broadly expressed actin nucleator, drives actin assembly during several intracellular stress response pathways. These newly described Arp2/3-mediated cytoskeletal rearrangements are coordinated by members of the Wiskott-Aldrich Syndrome Protein (WASP) family of actin nucleation-promoting factors. Thus, the Arp2/3 complex and WASP-family proteins are emerging as crucial players in cytoplasmic and nuclear activities including autophagy, apoptosis, chromatin dynamics, and DNA repair. Characterizations of the functions of the actin assembly machinery in such stress response mechanisms are advancing our understanding of both normal and pathogenic processes, and hold great promise for providing insights into organismal development and interventions for disease.
    Keywords DNA repair ; actin ; apoptosis ; autophagy ; chromatin ; eukaryotic cells ; microfilaments ; morphogenesis ; organelles ; stress response ; Arp2/3 ; ALS ; Cyto c ; DISC ; JMY ; MOMP ; N-WASP ; UPS ; WASP ; WASH ; WAVE ; WHAMM ; WHIMP ; Arp2/3 complex ; apoptosome ; caspase ; macropinocytosis ; mitochondria ; proteostasis ; transcription ; WASH complex
    Language English
    Dates of publication 2023-0302
    Size p. 151301
    Publishing place Elsevier GmbH
    Document type Article ; Online
    Note Pre-press version ; Use and reproduction
    ZDB-ID 391967-5
    ISSN 1618-1298 ; 0070-2463 ; 0171-9335
    ISSN (online) 1618-1298
    ISSN 0070-2463 ; 0171-9335
    DOI 10.1016/j.ejcb.2023.151301
    Database NAL-Catalogue (AGRICOLA)

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  9. Article ; Online: Genomic instability caused by Arp2/3 complex inactivation results in micronucleus biogenesis and cellular senescence.

    Haarer, Elena L / Theodore, Corey J / Guo, Shirley / Frier, Ryan B / Campellone, Kenneth G

    PLoS genetics

    2023  Volume 19, Issue 1, Page(s) e1010045

    Abstract: The Arp2/3 complex is an actin nucleator with well-characterized activities in cell morphogenesis and movement, but its roles in nuclear processes are relatively understudied. We investigated how the Arp2/3 complex affects genomic integrity and cell ... ...

    Abstract The Arp2/3 complex is an actin nucleator with well-characterized activities in cell morphogenesis and movement, but its roles in nuclear processes are relatively understudied. We investigated how the Arp2/3 complex affects genomic integrity and cell cycle progression using mouse fibroblasts containing an inducible knockout (iKO) of the ArpC2 subunit. We show that permanent Arp2/3 complex ablation results in DNA damage, the formation of cytosolic micronuclei, and cellular senescence. Micronuclei arise in ArpC2 iKO cells due to chromatin segregation defects during mitosis and premature mitotic exits. Such phenotypes are explained by the presence of damaged DNA fragments that fail to attach to the mitotic spindle, abnormalities in actin assembly during metaphase, and asymmetric microtubule architecture during anaphase. In the nuclei of Arp2/3-depleted cells, the tumor suppressor p53 is activated and the cell cycle inhibitor Cdkn1a/p21 mediates a G1 arrest. In the cytosol, micronuclei are recognized by the DNA sensor cGAS, which is important for stimulating a STING- and IRF3-associated interferon response. These studies establish functional requirements for the mammalian Arp2/3 complex in mitotic spindle organization and genome stability. They also expand our understanding of the mechanisms leading to senescence and suggest that cytoskeletal dysfunction is an underlying factor in biological aging.
    MeSH term(s) Animals ; Mice ; Actin-Related Protein 2-3 Complex/genetics ; Actin-Related Protein 2-3 Complex/metabolism ; Actins/metabolism ; Cellular Senescence/genetics ; DNA/metabolism ; Genomic Instability/genetics ; Mitosis/genetics
    Chemical Substances Actin-Related Protein 2-3 Complex ; Actins ; DNA (9007-49-2)
    Language English
    Publishing date 2023-01-27
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural
    ZDB-ID 2186725-2
    ISSN 1553-7404 ; 1553-7390
    ISSN (online) 1553-7404
    ISSN 1553-7390
    DOI 10.1371/journal.pgen.1010045
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  10. Article ; Online: Actin dynamics and function.

    Gupton, Stephanie L / Campellone, Kenneth G

    Molecular biology of the cell

    2018  Volume 29, Issue 6, Page(s) 696–697

    MeSH term(s) Actins/genetics ; Actins/metabolism ; Animals ; Congresses as Topic ; Humans ; Models, Biological ; Schizosaccharomyces
    Chemical Substances Actins
    Language English
    Publishing date 2018-03-11
    Publishing country United States
    Document type Journal Article
    ZDB-ID 1098979-1
    ISSN 1939-4586 ; 1059-1524
    ISSN (online) 1939-4586
    ISSN 1059-1524
    DOI 10.1091/mbc.E18-01-0010
    Database MEDical Literature Analysis and Retrieval System OnLINE

    Full text online

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    Zs.MO 410: Show issues

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