Actin Cytoskeletal Dynamics in Single-Cell Wound Repair

The plasma membrane protects the eukaryotic cell from its surroundings and is essential for cell viability; thus, it is crucial that membrane disruptions are repaired quickly to prevent immediate dyshomeostasis and cell death. Accordingly, cells have developed efficient repair mechanisms to rapidly reseal ruptures and reestablish membrane integrity. The cortical actin cytoskeleton plays an instrumental role in both plasma membrane resealing and restructuring in response to damage. Actin directly aids membrane repair or indirectly assists auxiliary repair mechanisms. Studies investigating single-cell wound repair have often focused on the recruitment and activation of specialized repair machinery, despite the undeniable need for rapid and dynamic cortical actin modulation; thus, the role of the cortical actin cytoskeleton during wound repair has received limited attention. This review aims to provide a comprehensive overview of membrane repair mechanisms directly or indirectly involving cortical actin cytoskeletal remodeling.

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Plasma membrane integrity in health and disease: significance and therapeutic potential

Cell Discovery ◽

10.1038/s41421-020-00233-2 ◽

2021 ◽

Vol 7 (1) ◽

Author(s):

Catarina Dias ◽

Jesper Nylandsted

Keyword(s):

Plasma Membrane ◽

Therapeutic Potential ◽

Calcium Influx ◽

Membrane Integrity ◽

Cell Types ◽

Physical Parameters ◽

Membrane Repair ◽

Plasma Membrane Integrity ◽

Repair Mechanisms ◽

And Function

AbstractMaintenance of plasma membrane integrity is essential for normal cell viability and function. Thus, robust membrane repair mechanisms have evolved to counteract the eminent threat of a torn plasma membrane. Different repair mechanisms and the bio-physical parameters required for efficient repair are now emerging from different research groups. However, less is known about when these mechanisms come into play. This review focuses on the existence of membrane disruptions and repair mechanisms in both physiological and pathological conditions, and across multiple cell types, albeit to different degrees. Fundamentally, irrespective of the source of membrane disruption, aberrant calcium influx is the common stimulus that activates the membrane repair response. Inadequate repair responses can tip the balance between physiology and pathology, highlighting the significance of plasma membrane integrity. For example, an over-activated repair response can promote cancer invasion, while the inability to efficiently repair membrane can drive neurodegeneration and muscular dystrophies. The interdisciplinary view explored here emphasises the widespread potential of targeting plasma membrane repair mechanisms for therapeutic purposes.

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Annexins in plasma membrane repair

Biological Chemistry ◽

10.1515/hsz-2016-0171 ◽

2016 ◽

Vol 397 (10) ◽

pp. 961-969 ◽

Cited By ~ 30

Author(s):

Theresa Louise Boye ◽

Jesper Nylandsted

Keyword(s):

Plasma Membrane ◽

Membrane Fusion ◽

Human Cancer ◽

Eukaryotic Cells ◽

Repair System ◽

Membrane Repair ◽

Cortical Actin ◽

Human Cancer Cells ◽

Repair Mechanisms ◽

Plasma Membrane Repair

Abstract Disruption of the plasma membrane poses deadly threat to eukaryotic cells and survival requires a rapid membrane repair system. Recent evidence reveal various plasma membrane repair mechanisms, which are required for cells to cope with membrane lesions including membrane fusion and replacement strategies, remodeling of cortical actin cytoskeleton and vesicle wound patching. Members of the annexin protein family, which are Ca2+-triggered phospholipid-binding proteins emerge as important components of the plasma membrane repair system. Here, we discuss the mechanisms of plasma membrane repair involving annexins spanning from yeast to human cancer cells.

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Ultrastructure of the yeast actin cytoskeleton and its association with the plasma membrane.

The Journal of Cell Biology ◽

10.1083/jcb.125.2.381 ◽

1994 ◽

Vol 125 (2) ◽

pp. 381-391 ◽

Cited By ~ 245

Author(s):

J Mulholland ◽

D Preuss ◽

A Moon ◽

A Wong ◽

D Drubin ◽

...

Keyword(s):

Plasma Membrane ◽

Actin Cytoskeleton ◽

Binding Proteins ◽

Ultrastructural Level ◽

Actin Binding ◽

Cortical Actin ◽

Actin Binding Proteins ◽

Double Labeling ◽

Wall Growth ◽

Cortical Cytoskeleton

We characterized the yeast actin cytoskeleton at the ultrastructural level using immunoelectron microscopy. Anti-actin antibodies primarily labeled dense, patchlike cortical structures and cytoplasmic cables. This localization recapitulates results obtained with immunofluorescence light microscopy, but at much higher resolution. Immuno-EM double-labeling experiments were conducted with antibodies to actin together with antibodies to the actin binding proteins Abp1p and cofilin. As expected from immunofluorescence experiments, Abp1p, cofilin, and actin colocalized in immuno-EM to the dense patchlike structures but not to the cables. In this way, we can unambiguously identify the patches as the cortical actin cytoskeleton. The cortical actin patches were observed to be associated with the cell surface via an invagination of plasma membrane. This novel cortical cytoskeleton-plasma membrane interface appears to consist of a fingerlike invagination of plasma membrane around which actin filaments and actin binding proteins are organized. We propose a possible role for this unique cortical structure in wall growth and osmotic regulation.

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Cell wound repair in Drosophila occurs through three distinct phases of membrane and cytoskeletal remodeling

The Journal of Cell Biology ◽

10.1083/jcb.201011018 ◽

2011 ◽

Vol 193 (3) ◽

pp. 455-464 ◽

Cited By ~ 78

Author(s):

Maria Teresa Abreu-Blanco ◽

Jeffrey M. Verboon ◽

Susan M. Parkhurst

Keyword(s):

Cell Death ◽

Plasma Membrane ◽

Single Cell ◽

Wound Repair ◽

Single Cells ◽

Repair Process ◽

Wound Edge ◽

Cytoskeletal Remodeling ◽

Tissue Integrity ◽

E Cadherin

When single cells or tissues are injured, the wound must be repaired quickly in order to prevent cell death, loss of tissue integrity, and invasion by microorganisms. We describe Drosophila as a genetically tractable model to dissect the mechanisms of single-cell wound repair. By analyzing the expression and the effects of perturbations of actin, myosin, microtubules, E-cadherin, and the plasma membrane, we define three distinct phases in the repair process—expansion, contraction, and closure—and identify specific components required during each phase. Specifically, plasma membrane mobilization and assembly of a contractile actomyosin ring are required for this process. In addition, E-cadherin accumulates at the wound edge, and wound expansion is excessive in E-cadherin mutants, suggesting a role for E-cadherin in anchoring the actomyosin ring to the plasma membrane. Our results show that single-cell wound repair requires specific spatial and temporal cytoskeleton responses with distinct components and mechanisms required at different stages of the process.

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Endogenous plasma membrane t-SNARE syntaxin 4 is present in rab11 positive endosomal membranes and associates with cortical actin cytoskeleton

FEBS Letters ◽

10.1016/s0014-5793(02)03605-0 ◽

2002 ◽

Vol 531 (3) ◽

pp. 513-519 ◽

Cited By ~ 30

Author(s):

Arja M Band ◽

Heidi Ali ◽

Maria K Vartiainen ◽

Saara Welti ◽

Pekka Lappalainen ◽

...

Keyword(s):

Plasma Membrane ◽

Actin Cytoskeleton ◽

Cortical Actin ◽

Endosomal Membranes ◽

Syntaxin 4

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Exocytosis of acid sphingomyelinase by wounded cells promotes endocytosis and plasma membrane repair

The Journal of Cell Biology ◽

10.1083/jcb.201003053 ◽

2010 ◽

Vol 189 (6) ◽

pp. 1027-1038 ◽

Cited By ~ 197

Author(s):

Christina Tam ◽

Vincent Idone ◽

Cecilia Devlin ◽

Maria Cecilia Fernandes ◽

Andrew Flannery ◽

...

Keyword(s):

Plasma Membrane ◽

Lysosomal Enzyme ◽

Membrane Integrity ◽

Acid Sphingomyelinase ◽

Membrane Repair ◽

Membrane Injury ◽

Cellular Survival ◽

Plasma Membrane Repair ◽

Membrane Resealing ◽

Niemann Pick

Rapid plasma membrane resealing is essential for cellular survival. Earlier studies showed that plasma membrane repair requires Ca2+-dependent exocytosis of lysosomes and a rapid form of endocytosis that removes membrane lesions. However, the functional relationship between lysosomal exocytosis and the rapid endocytosis that follows membrane injury is unknown. In this study, we show that the lysosomal enzyme acid sphingomyelinase (ASM) is released extracellularly when cells are wounded in the presence of Ca2+. ASM-deficient cells, including human cells from Niemann-Pick type A (NPA) patients, undergo lysosomal exocytosis after wounding but are defective in injury-dependent endocytosis and plasma membrane repair. Exogenously added recombinant human ASM restores endocytosis and resealing in ASM-depleted cells, suggesting that conversion of plasma membrane sphingomyelin to ceramide by this lysosomal enzyme promotes lesion internalization. These findings reveal a molecular mechanism for restoration of plasma membrane integrity through exocytosis of lysosomes and identify defective plasma membrane repair as a possible component of the severe pathology observed in NPA patients.

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Lysosomal retargeting of Myoferlin mitigates membrane stress to enable pancreatic cancer growth

10.1101/2021.01.04.425106 ◽

2021 ◽

Author(s):

Suprit Gupta ◽

Julian Yano ◽

Htet Htwe Htwe ◽

Hijai R. Shin ◽

Zeynep Cakir ◽

...

Keyword(s):

Pancreatic Cancer ◽

Plasma Membrane ◽

Membrane Damage ◽

Membrane Integrity ◽

Human Pancreatic Cancer ◽

Cancer Growth ◽

Membrane Repair ◽

Pancreatic Cancer Cells ◽

Plasma Membrane Repair

AbstractLysosomes must maintain integrity of their limiting membrane to ensure efficient fusion with incoming organelles and degradation of substrates within their lumen. Pancreatic cancer cells upregulate lysosomal biogenesis to enhance nutrient recycling and stress resistance, but whether dedicated programs for maintaining lysosomal membrane integrity facilitate pancreatic cancer growth is unknown. Using proteomic-based organelle profiling, we identify the Ferlin family plasma membrane repair factor, Myoferlin, as selectively and highly enriched on the membrane of pancreatic cancer lysosomes. Mechanistically, lysosome localization of Myoferlin is necessary and sufficient for maintenance of lysosome health and provides an early-acting protective system against membrane damage that is independent from the endosomal sorting complex required for transport (ESCRT)-mediated repair network. Myoferlin is upregulated in human pancreatic cancer, predicts poor survival, and its ablation severely impairs lysosome function and tumour growth in vivo. Thus, retargeting of plasma membrane repair factors enhances pro-oncogenic activities of the lysosome.

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Dealing with damage: Plasma membrane repair mechanisms

Biochimie ◽

10.1016/j.biochi.2014.08.008 ◽

2014 ◽

Vol 107 ◽

pp. 66-72 ◽

Cited By ~ 39

Author(s):

Annette Draeger ◽

Roman Schoenauer ◽

Alexander P. Atanassoff ◽

Heidi Wolfmeier ◽

Eduard B. Babiychuk

Keyword(s):

Plasma Membrane ◽

Membrane Repair ◽

Repair Mechanisms ◽

Plasma Membrane Repair

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Perforin activates clathrin- and dynamin-dependent endocytosis, which is required for plasma membrane repair and delivery of granzyme B for granzyme-mediated apoptosis

Blood ◽

10.1182/blood-2009-10-246116 ◽

2010 ◽

Vol 115 (8) ◽

pp. 1582-1593 ◽

Cited By ~ 74

Author(s):

Jerome Thiery ◽

Dennis Keefe ◽

Saviz Saffarian ◽

Denis Martinvalet ◽

Michael Walch ◽

...

Keyword(s):

Plasma Membrane ◽

Cytotoxic T Lymphocytes ◽

Granzyme B ◽

Membrane Integrity ◽

Effector Proteins ◽

Target Cells ◽

Membrane Repair ◽

Early Endosomes ◽

Plasma Membrane Repair ◽

Immunologic Synapse

Abstract Cytotoxic T lymphocytes and natural killer cells destroy target cells via the polarized exocytosis of lytic effector proteins, perforin and granzymes, into the immunologic synapse. How these molecules enter target cells is not fully understood. It is debated whether granzymes enter via perforin pores formed at the plasma membrane or whether perforin and granzymes are first endocytosed and granzymes are then released from endosomes into the cytoplasm. We previously showed that perforin disruption of the plasma membrane induces a transient Ca2+ flux into the target cell that triggers a wounded membrane repair response in which lysosomes and endosomes donate their membranes to reseal the damaged membrane. Here we show that perforin activates clathrin- and dynamin-dependent endocytosis, which removes perforin and granzymes from the plasma membrane to early endosomes, preserving outer membrane integrity. Inhibiting clathrin- or dynamin-dependent endocytosis shifts death by perforin and granzyme B from apoptosis to necrosis. Thus by activating endocytosis to preserve membrane integrity, perforin facilitates granzyme uptake and avoids the proinflammatory necrotic death of a membrane-damaged cell.

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