Functional and Genetic Analysis of Plectin in Skin and Muscle

Rezniczek, Günther A.; Winter, Lilli; Walko, Gernot; Wiche, Gerhard

doi:10.1016/bs.mie.2015.05.003

Cited by 18 publications

(19 citation statements)

References 44 publications

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“…For direct lysis, confluent endothelial cell cultures, washed twice with phosphate-buffered saline (PBS), were overlaid with 50 mM Tris-HCl, pH 6.8, 100 mM DTT, 2% SDS, 0.1% Bromophenol Blue and 10% glycerol. Detergent extraction of soluble vimentin was performed as described previously ( Rezniczek et al, 2015 ). Aliquots of total cell lysates or cell fractions were subjected to SDS-PAGE and, after immunoblotting using peroxidase-coupled secondary antibodies, protein bands were visualized by exposure to X-ray film ( Osmanagic-Myers and Wiche, 2004 ).…”

Section: Methodsmentioning

confidence: 99%

Plectin reinforces vascular integrity by mediating vimentin-actin network crosstalk

Osmanagic‐Myers

Rus

Wolfram

et al. 2015

Journal of Cell Science

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Mutations in the cytoskeletal linker protein plectin result in multisystemic diseases affecting skin and muscle with indications of additional vascular system involvement. To study the mechanisms underlying vascular disorders, we established plectin-deficient endothelial cell and mouse models. We show that apart from perturbing the vimentin cytoskeleton of endothelial cells, plectin deficiency leads to severe distortions of adherens junctions (AJs), as well as tight junctions, accompanied by an upregulation of actin stress fibres and increased cellular contractility. Plectin-deficient endothelial cell layers were more leaky and showed reduced mechanical resilience in fluid-shear stress and mechanical stretch experiments. We suggest that the distorted AJs and upregulated actin stress fibres in plectin-deficient cells are rooted in perturbations of the vimentin cytoskeleton, as similar phenotypes could be mimicked in wild-type cells by disruption of vimentin filaments. In vivo studies in endothelium-restricted conditional plectin-knockout mice revealed significant distortions of AJs in stress-prone aortic arch regions and increased pulmonary vascular leakage. Our study opens a new perspective on cytoskeleton-controlled vascular permeability, where a plectin-organized vimentin scaffold keeps actomyosin contractility ‘in-check’ and maintains AJ homeostasis.

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Section: Methodsmentioning

confidence: 99%

Plectin reinforces vascular integrity by mediating vimentin-actin network crosstalk

Osmanagic‐Myers

Rus

Wolfram

et al. 2015

Journal of Cell Science

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“…PLEC encodes plectin, a large cytoskeletal protein that regulates signalling from the extracellular environment to the cell nucleus 18 , 19 . Plectin enables cells to respond to external mechanical stimuli and forces, such as those experienced by the chondrocyte during joint movement 20 .…”

Section: Introductionmentioning

confidence: 99%

Multi-tissue epigenetic analysis of the osteoarthritis susceptibility locus mapping to the plectin gene PLEC

Sorial

Hofer

Tselepi

et al. 2020

Osteoarthritis and Cartilage

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Summary Objective In cartilage, the osteoarthritis (OA) associated single nucleotide polymorphism (SNP) rs11780978 correlates with differential expression of PLEC , and with differential methylation of PLEC CpG dinucleotides, forming eQTLs and mQTLs respectively. This implies that methylation links chondrocyte genotype and phenotype, thus driving the functional effect of this genetic risk signal. PLEC encodes plectin, a cytoskeletal protein that enables tissues to respond to mechanical forces. We sought to assess whether these PLEC functional effects were cartilage specific. Method Cartilage, fat pad, synovium and peripheral blood were collected from patients undergoing arthroplasty. PLEC CpGs were analysed for mQTLs and allelic expression imbalance (AEI) was performed to test for eQTLs. Plectin was knocked down in a mesenchymal stem cell (MSC) line using CRISPR/Cas9 and cells phenotyped by RNA-sequencing. Results mQTLs were discovered in fat pad, synovium and blood. Their effects were however stronger in the joint tissues and of comparable effect between these tissues. We observed AEI in synovium in the same direction as for cartilage and correlations between methylation and PLEC expression. Knocking-down plectin impacted on pathways reported to have a role in OA, including Wnt signalling, glycosaminoglycan biosynthesis and immune regulation. Conclusions Synovium is also a target of the rs11780978 OA association functionally operating on PLEC . In fat pad, mQTLs were identified but these did not correlate with PLEC expression, suggesting the functional effect is not joint-wide. Our study highlights interplay between genetic risk, DNA methylation and gene expression in OA, and reveals clear differences between tissues from the same diseased joint.

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“…Recently reported mutations of the plectin gene within exons 1f ( 61 ) and 1a ( 62 ), causing muscle-only limb girdle muscular dystrophy (MD) and skin-only epidermolysis bullosa simplex (EBS), respectively, as well as the analyses of diverse plectin-isoform-deficient mouse lines ( 63 ) clearly showed that the different isoforms of plectin are responsible for distinct functions. The fact that P1 is involved in proper nuclear architecture and function opens a new perspective on the role of cytolinkers in mechanotransduction.…”

Section: Discussionmentioning

confidence: 99%

Plectin isoform 1-dependent nuclear docking of desmin networks affects myonuclear architecture and expression of mechanotransducers

2015

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Plectin is a highly versatile cytoskeletal protein that acts as a mechanical linker between intermediate filament (IF) networks and various cellular structures. The protein is crucial for myofiber integrity. Its deficiency leads to severe pathological changes in skeletal muscle fibers of patients suffering from epidermolysis bullosa simplex with muscular dystrophy (EBS-MD). Skeletal muscle fibers express four major isoforms of plectin which are distinguished solely by alternative, relatively short, first exon-encoded N-terminal sequences. Each one of these isoforms is localized to a different subcellular compartment and plays a specific role in maintaining integrity and proper function(s) of myofibers. The unique role of individual isoforms is supported by distinct phenotypes of isoform-specific knockout mice and recently discovered mutations in first coding exons of plectin that lead to distinct, tissue-specific, pathological abnormalities in humans. In this study, we demonstrate that the lack of plectin isoform 1 (P1) in myofibers of mice leads to alterations of nuclear morphology, similar to those observed in various forms of MD. We show that P1-mediated targeting of desmin IFs to myonuclei is essential for maintenance of their typically spheroidal architecture as well as their proper positioning and movement along the myofiber. Furthermore, we show that P1 deficiency affects chromatin modifications and the expression of genes involved in various cellular functions, including signaling pathways mediating mechanotransduction. Mechanistically, P1 is shown to specifically interact with the myonuclear membrane-associated (BAR domain-containing) protein endophilin B. Our results open a new perspective on cytoskeleton-nuclear crosstalk via specific cytolinker proteins.

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Functional and Genetic Analysis of Plectin in Skin and Muscle

Cited by 18 publications

References 44 publications

Plectin reinforces vascular integrity by mediating vimentin-actin network crosstalk

Plectin reinforces vascular integrity by mediating vimentin-actin network crosstalk

Multi-tissue epigenetic analysis of the osteoarthritis susceptibility locus mapping to the plectin gene PLEC

Plectin isoform 1-dependent nuclear docking of desmin networks affects myonuclear architecture and expression of mechanotransducers

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