Mechanical Stabilization of the Glandular Acinus by Linker of Nucleoskeleton and Cytoskeleton Complex

Zhang, Qiao; Narayanan, Vani; Mui, Keeley L.; O’Bryan, Christopher S.; Anderson, Ruthellen H.; Kc, Birendra; Cabe, Jolene I; Denis, Kevin; Antoku, Susumu; Roux, Kyle J.; Dickinson, Richard B.; Angelini, Thomas E.; Gundersen, Gregg G.; Conway, Daniel E.; Lele, Tanmay P.

doi:10.1016/j.cub.2019.07.021

Cited by 26 publications

(33 citation statements)

References 87 publications

(123 reference statements)

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“…To understand if this change in nuclear shape in response to SMG depends on the LINC complex, we used a doxycycline inducible MCF 10A cell line where the LINC complex function is disrupted. This inducible cell line stably expresses SS-HA-SUN1L-KDEL (signal sequence-hemaglutinin epitope tagged to SUN1 protein, with a lumenal domain-ER retrieval amino acid sequence; SUN1L-KDEL) [ 25 ]. SUN1L-KDEL acts in a dominant negative fashion disrupting the LINC complex by interfering with the connection between the endogenous KASH domain of nesprin-2 and nesprin-4 proteins and the SUN1 proteins in the nuclear envelope [ 3 , 21 , 25 ].…”

Section: Resultsmentioning

confidence: 99%

“…This inducible cell line stably expresses SS-HA-SUN1L-KDEL (signal sequence-hemaglutinin epitope tagged to SUN1 protein, with a lumenal domain-ER retrieval amino acid sequence; SUN1L-KDEL) [ 25 ]. SUN1L-KDEL acts in a dominant negative fashion disrupting the LINC complex by interfering with the connection between the endogenous KASH domain of nesprin-2 and nesprin-4 proteins and the SUN1 proteins in the nuclear envelope [ 3 , 21 , 25 ]. Cells expressing a similar doxycycline inducible construct SS-GFP-KDEL (GFP-KDEL) were used as the corresponding paired control.…”

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

“…Human breast epithelial cells (MCF 10A from ATCC), MCF 10A SS-GFP-KDEL and MCF 10A SS-GFP-SUN1L-KDEL cells (created by Dr. Tanmay Lele and Dr. Kyle Roux [ 21 , 25 ]) were a kind gift to us. The inducible cell lines were created with MCF 10A TET-ON (Clontech) transduced retrovirally with pRetroX-Tight.puro containing either SS-HA-GFP-SUN1L-KDEL for LINC disruption or SS-HA-GFP-KDEL for controls [ 25 ]. To create a stable line, cell selection was performed with 0.5 μg/mL puromycin and then screened by immunofluorescence postinduction with or without 1 μg/mL doxycycline for 18 h. The cells were cultured in DMEM/F12 (Thermofisher, Grand Island, NY, USA) without phenol red, supplemented with 5% ( w/v ) horse serum, cholera toxin (Sigma, Saint Louis, MO USA), epidermal growth factor (Peprotech, Cranbury, NJ, USA), hydrocortisone (Sigma), insulin (Sigma) and penicillin streptomycin (Thermofisher, Grand Island, NY, USA).…”

Section: Methodsmentioning

confidence: 99%

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Changes in Nuclear Shape and Gene Expression in Response to Simulated Microgravity Are LINC Complex-Dependent

Neelam

Richardson

Barker

et al. 2020

IJMS

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Microgravity is known to affect the organization of the cytoskeleton, cell and nuclear morphology and to elicit differential expression of genes associated with the cytoskeleton, focal adhesions and the extracellular matrix. Although the nucleus is mechanically connected to the cytoskeleton through the Linker of Nucleoskeleton and Cytoskeleton (LINC) complex, the role of this group of proteins in these responses to microgravity has yet to be defined. In our study, we used a simulated microgravity device, a 3-D clinostat (Gravite), to investigate whether the LINC complex mediates cellular responses to the simulated microgravity environment. We show that nuclear shape and differential gene expression are both responsive to simulated microgravity in a LINC-dependent manner and that this response changes with the duration of exposure to simulated microgravity. These LINC-dependent genes likely represent elements normally regulated by the mechanical forces imposed by gravity on Earth.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Changes in Nuclear Shape and Gene Expression in Response to Simulated Microgravity Are LINC Complex-Dependent

Neelam

Richardson

Barker

et al. 2020

IJMS

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“…This structure has been shown to regulate nuclear movement and positioning (Luxton et al, 2010;Petrie et al, 2012), DNA repair (Lei et al, 2012;Lottersberger et al, 2015), and transmission of mechanical forces from the cell surface to the nucleus (Chambliss et al, 2013;Tajik et al, 2016;Mayer et al, 2019). More recently our group showed that the LINC complex is important in the mechanical stability of epithelial glandular acini (Zhang et al, 2019), suggesting that the LINC complex also has an important role in the mechanical stability of multicellular tissues.…”

Section: Introductionmentioning

confidence: 97%

The LINC complex is required for endothelial cell adhesion and adaptation to shear stress and cyclic stretch

Denis

Cabe

Danielsson

et al. 2021

MBoC

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The Linker of Nucleoskeleton and Cytoskeleton (LINC) complex is a structure consisting of nesprin, SUN, and lamin proteins. A principal function of the LINC complex is anchoring the nucleus to the actin, microtubule, and intermediate filament cytoskeletons. The LINC complex is present in nearly all cell types, including endothelial cells. Endothelial cells line the inner most surfaces of blood vessels, and are critical for blood vessel barrier function. In addition, endothelial cells have specialized functions, including adaptation to the mechanical forces of blood flow. Previous studies have shown that depletion of individual nesprin isoforms results in impaired endothelial cell function. To further investigate the role of the LINC complex in endothelial cells we utilized dominant negative KASH (DN-KASH), a dominant negative protein which displaces endogenous nesprins from the nuclear envelope and disrupts nuclear-cytoskeletal connections. Endothelial cells expressing DN-KASH had altered cell-cell adhesion and barrier function, as well as altered cell-matrix adhesion and focal adhesion dynamics. In addition, cells expressing DN-KASH failed to properly adapt to shear stress or cyclic stretch. DN-KASH expressing cells exhibited impaired collective cell migration in wound healing and angiogenesis assays. Our results demonstrate the importance of an intact LINC complex in endothelial cell function and homeostasis. [Media: see text] [Media: see text] [Media: see text] [Media: see text] [Media: see text] [Media: see text] [Media: see text] [Media: see text] [Media: see text] [Media: see text]

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Osteoprotegerin is sensitive to actomyosin tension in human periodontal ligament fibroblasts

Tamashunas

Katiyar

Zhang

et al. 2021

Journal Cellular Physiology

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Periodontal ligament fibroblasts (PdLFs) are an elongated cell type in the periodontium with matrix and bone regulatory functions which become abnormal in periodontal disease (PD). Here we found that the normally elongated and oriented PdLF nucleus becomes rounded and loses orientation in a mouse model of PD. Using in vitro micropatterning of cultured primary PdLF cell shape, we show that PdLF elongation correlates with nuclear elongation and the presence of thicker, contractile F‐actin fibers. The rounded nuclei in mouse PD models in vivo are, therefore, indicative of reduced actomyosin tension. Inhibiting actomyosin contractility by inhibiting myosin light chain kinase, Rho kinase or myosin ATPase activity, in cultured PdLFs each consistently reduced messenger RNA levels of bone regulatory protein osteoprotegerin (OPG). Infection of cultured PdLFs with two different types of periodontal bacteria (Porphyromonas gingivalis and Fusobacterium nucleatum) failed to recapitulate the observed nuclear rounding in vivo, upregulated nonmuscle myosin II phosphorylation and downregulated OPG. Collectively, our results add support to the hypothesis that PdLF contractility becomes decreased and contributes to disease progression in PD.

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Mechanical Stabilization of the Glandular Acinus by Linker of Nucleoskeleton and Cytoskeleton Complex

Cited by 26 publications

References 87 publications

Changes in Nuclear Shape and Gene Expression in Response to Simulated Microgravity Are LINC Complex-Dependent

Changes in Nuclear Shape and Gene Expression in Response to Simulated Microgravity Are LINC Complex-Dependent

The LINC complex is required for endothelial cell adhesion and adaptation to shear stress and cyclic stretch

Osteoprotegerin is sensitive to actomyosin tension in human periodontal ligament fibroblasts

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