Myosin IIA Dependent Retrograde Flow Drives 3D Cell Migration

Abstract The immune response relies on the migration of leukocytes and on their ability to stop in precise anatomical locations to fulfil their task. How leukocyte migration and function are coordinated is unknown. Here we show that in immature dendritic cells, which patrol their environment by engulfing extracellular material, cell migration and antigen capture are antagonistic. This antagonism results from transient enrichment of myosin IIA at the cell front, which disrupts the back-to-front gradient of the motor protein, slowing down locomotion but promoting antigen capture. We further highlight that myosin IIA enrichment at the cell front requires the MHC class II-associated invariant chain (Ii). Thus, by controlling myosin IIA localization, Ii imposes on dendritic cells an intermittent antigen capture behaviour that might facilitate environment patrolling. We propose that the requirement for myosin II in both cell migration and specific cell functions may provide a general mechanism for their coordination in time and space.

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Nucleus and nucleus-cytoskeleton connections in 3D cell migration

Experimental Cell Research ◽

10.1016/j.yexcr.2016.09.001 ◽

2016 ◽

Vol 348 (1) ◽

pp. 56-65 ◽

Cited By ~ 19

Author(s):

Lingling Liu ◽

Qing Luo ◽

Jinghui Sun ◽

Guanbin Song

Keyword(s):

Cell Migration ◽

3D Cell Migration

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Retrograde Flow for Forward Cell Migration

Science Signaling ◽

10.1126/scisignal.2005714 ◽

2014 ◽

Vol 7 (335) ◽

pp. ec194-ec194

Author(s):

W. Wong

Keyword(s):

Cell Migration ◽

Retrograde Flow

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ARID1A interacts with nonmuscle myosin IIA to regulate cancer cell motility.

Journal of Clinical Oncology ◽

10.1200/jco.2019.37.15_suppl.e17036 ◽

2019 ◽

Vol 37 (15_suppl) ◽

pp. e17036-e17036

Author(s):

Oloruntoba Ismail Osagie ◽

Zhigui Li ◽

Shijun Mi ◽

Jennifer T Aguilan ◽

Gloria S. Huang

Keyword(s):

Cell Migration ◽

Cell Motility ◽

Cell Lines ◽

Cancer Cell ◽

Protein Interaction ◽

Protein Complex ◽

Protein Identification ◽

Cancer Cell Motility ◽

Protein Protein Interaction ◽

Myosin Iia

e17036 Background: ARID1A (BAF250A), a member of the SWI/SNF chromatin remodeling complex, is one of the most frequently mutated genes in human cancer. Here we report the discovery of a novel protein-protein interaction between ARID1A and the actin-binding motor protein, non-muscle myosin IIA (NM IIA) encoded by the myosin heavy chain 9 ( MYH9). Methods: The ARID1A immunoprecipitated protein complex was separated by gel electrophoresis followed by analysis of the peptide digested gel bands by C18-Reversed Phase chromatography using an Ultimate 3000 RSLCnano System (Thermo Scientific) equipped with an Acclaim PepMap C18 column (Thermo Scientific) and connected to a TriVersa NanoMate nanoelectrospray source (Advion) and a linear ion trap LTQ-XL mass spectrometer (Thermo Scientific). Protein identification was performed by Mascot search engine v. 2.5.1 (Matrix Science) against NCBI Homo sapiens database. Scaffold software v. 4.5.1 (Proteome Software Inc.) was used to validate the MS/MS peptide and protein identification based on 99% protein and 95% peptide probabilities. Immunoprecipitation and immunoblotting were done to evaluate the protein-protein interaction in ARID1A-wild type cell lines. Isogenic engineered cell lines, ES2 shRNA-control or shRNA- ARID1A stable transfection , and HCT116 control or ARID1A knockout by CRISPR-Cas9 (Horizon Discovery) were used to evaluate the effect of ARID1A loss on NM IIA expression and phosphorylation, and on cell migration by in vitro scratch assay with time lapse imaging. Results: Scaffold analysis of peptide spectra identified NM IIA with > 99% probability in the ARID1A immunopurified protein complex. In the ARID1A wildtype cell lines ES2 and KLE, endogenous NM IIA co-immunoprecipitated with ARID1A and vice versa. ES2 sh ARID1A cells had decreased total and phosphorylated NM IIA expression, and impaired cell migration compared to control cells. Similarly, HCT116 ARID1A homozygous knockout cells had impaired cell migration compared with HCT116 control cells. Conclusions: We report for the first time that ARID1A interacts with NM IIA to regulate cancer cell motility. Further investigation is ongoing to elucidate the significance of this newly identified function of ARID1A.

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Nonpolarized signaling reveals two distinct modes of 3D cell migration

The Journal of Cell Biology ◽

10.1083/jcb.201201124 ◽

2012 ◽

Vol 197 (3) ◽

pp. 439-455 ◽

Cited By ~ 232

Author(s):

Ryan J. Petrie ◽

Núria Gavara ◽

Richard S. Chadwick ◽

Kenneth M. Yamada

Keyword(s):

Cell Migration ◽

Intracellular Signaling ◽

Myosin Ii ◽

Three Dimensional ◽

Leading Edge ◽

Elastic Behavior ◽

Collagen Gels ◽

Primary Fibroblasts ◽

Context Specific ◽

3D Cell Migration

We search in this paper for context-specific modes of three-dimensional (3D) cell migration using imaging for phosphatidylinositol (3,4,5)-trisphosphate (PIP3) and active Rac1 and Cdc42 in primary fibroblasts migrating within different 3D environments. In 3D collagen, PIP3 and active Rac1 and Cdc42 were targeted to the leading edge, consistent with lamellipodia-based migration. In contrast, elongated cells migrating inside dermal explants and the cell-derived matrix (CDM) formed blunt, cylindrical protrusions, termed lobopodia, and Rac1, Cdc42, and PIP3 signaling was nonpolarized. Reducing RhoA, Rho-associated protein kinase (ROCK), or myosin II activity switched the cells to lamellipodia-based 3D migration. These modes of 3D migration were regulated by matrix physical properties. Specifically, experimentally modifying the elasticity of the CDM or collagen gels established that nonlinear elasticity supported lamellipodia-based migration, whereas linear elasticity switched cells to lobopodia-based migration. Thus, the relative polarization of intracellular signaling identifies two distinct modes of 3D cell migration governed intrinsically by RhoA, ROCK, and myosin II and extrinsically by the elastic behavior of the 3D extracellular matrix.

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