ADF and cofilin-1 collaborate to promote cortical actin flow and the leader bleb-based migration of confined cells

ADF and cofilin-1 collaborate to promote cortical actin flow and the leader bleb-based migration of confined cells

Publication date: Jan 11, 2021

Melanoma cells have been shown to undergo fast amoeboid or Leader Bleb-Based Migration (LBBM), requiring a large and stable bleb for migration. In leader blebs, is a rapid flow of cortical actin that drives the cell forward. Here, we tested the hypothesis that the actin severing factors, ADF and cofilin-1, are essential for contractility and actin flow for LBBM. Using RNAi in melanoma A375 cells, we find that co-depleting ADF and cofilin-1 led to a large increase in the level of cortical actin, suggesting that ADF and cofilin-1, together, regulate cortical actin in these cells. Moreover, RNAi of ADF and cofilin-1 increased the number of cortical, polymerization competent, barbed-ends. Therefore, severing by these proteins appears to promote cortical actin turnover. Furthermore, actin severing factors can promote contractility through the regulation of actin architecture. In agreement with this concept, RNAi of ADF and cofilin-1 led to a significant decrease in cell stiffness. As LBBM is stimulated by cell confinement, we then used confined cells to evaluate the role of ADF and cofilin-1 in regulating actin dynamics. We found cofilin-1 to be enriched at leader bleb necks, whereas RNAi of ADF and cofilin-1 reduced bleb sizes and the frequency of motile cells. Strikingly, cells without ADF and cofilin-1 had blebs with abnormally long necks. Many blebs failed to retract and displayed slower actin turnover and flow in the absence of ADF and cofilin-1. Collectively, our data identifies ADF and cofilin-1 as actin remodeling factors required for the amoeboid migration of melanoma cells.

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Concepts Keywords
Actin Serum free media
Albany Min media
Albany College High resolution imaging
Amc Min imaging
Amoeboid Oil objective
Assembly End assay protocol
Cofilin Plates complete media
Cortical Online culture transfection
Cytoskeleton Immunofluorescence imaging
Filament Branches of biology
Force Structural proteins
Frequency Cell biology
Friction Cytoskeleton
Intracellular ADF/Cofilin family
Lamellipodia Cofilin 1
Mail Microfilament
Melanoma Actin
Mesenchymal Cell cortex
Metastasis Lim kinase
Motile Bleb
Myosin Invasion
Perpetuity Antibodies
Phenotypic Polymerization
Phosphorylation Laser
Physicochemical 3G
Plasticity End assay protocol
Polymerization
Retrograde
RNAi
Scotland
Stiffness

Semantics

Type Source Name
disease MESH bleb
disease MESH Melanoma
pathway KEGG Melanoma
disease MESH cancer
disease MESH metastasis
disease MESH separation
drug DRUGBANK Serine
disease MESH lung adenocarcinoma
disease MESH defects
drug DRUGBANK Cefaclor
drug DRUGBANK Dextrose unspecified form
disease MESH biopsy
drug DRUGBANK Latrunculin A
drug DRUGBANK Dimethyl sulfoxide
drug DRUGBANK Edetic Acid
drug DRUGBANK Sodium lauryl sulfate
drug DRUGBANK Deoxycholic Acid
drug DRUGBANK Tromethamine
drug DRUGBANK Flunarizine
drug DRUGBANK Medical air
drug DRUGBANK Esomeprazole
drug DRUGBANK Aspartame
drug DRUGBANK L-Lysine
drug DRUGBANK Potassium Chloride
drug DRUGBANK ATP
drug DRUGBANK Gelatin
drug DRUGBANK Adenosine 5′-phosphosulfate
drug DRUGBANK Water
drug DRUGBANK Dacarbazine
drug DRUGBANK Hyaluronic acid
drug DRUGBANK Etoperidone

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