title: Stimulation of cortical myosin phosphorylation by p114RhoGEF drives cell migration and tumor cell invasion.
abstract: Actinomyosin activity is an important driver of cell locomotion and has been shown to promote collective cell migration of epithelial sheets as well as single cell migration and tumor cell invasion. However, the molecular mechanisms underlying activation of cortical myosin to stimulate single cell movement, and the relationship between the mechanisms that drive single cell locomotion and those that mediate collective cell migration of epithelial sheets are incompletely understood. Here, we demonstrate that p114RhoGEF, an activator of RhoA that associates with non-muscle myosin IIA, regulates collective cell migration of epithelial sheets and tumor cell invasion. Depletion of p114RhoGEF resulted in specific spatial inhibition of myosin activation at cell-cell contacts in migrating epithelial sheets and the cortex of migrating single cells, but only affected double and not single phosphorylation of myosin light chain. In agreement, overall elasticity and contractility of the cells, processes that rely on persistent and more constant forces, were not affected, suggesting that p114RhoGEF mediates process-specific myosin activation. Locomotion was p114RhoGEF-dependent on Matrigel, which favors more roundish cells and amoeboid-like actinomyosin-driven movement, but not on fibronectin, which stimulates flatter cells and lamellipodia-driven, mesenchymal-like migration. Accordingly, depletion of p114RhoGEF led to reduced RhoA, but increased Rac activity. Invasion of 3D matrices was p114RhoGEF-dependent under conditions that do not require metalloproteinase activity, supporting a role of p114RhoGEF in myosin-dependent, amoeboid-like locomotion. Our data demonstrate that p114RhoGEF drives cortical myosin activation by stimulating myosin light chain double phosphorylation and, thereby, collective cell migration of epithelial sheets and amoeboid-like motility of tumor cells.
authors.affiliation: Department of Cell Biology, UCL Institute of Ophthalmology, University College London, London, United Kingdom.
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Terry
authors.forename: Ahmed
authors.initials: A
authors.lastname: Elbediwy
authors.forename: Ceniz
authors.initials: C
authors.lastname: Zihni
authors.forename: Andrew R
authors.initials: AR
authors.lastname: Harris
authors.forename: Maryse
authors.initials: M
authors.lastname: Bailly
authors.forename: Guillaume T
authors.initials: GT
authors.lastname: Charras
authors.forename: Maria S
authors.initials: MS
authors.lastname: Balda
authors.forename: Karl
authors.initials: K
authors.lastname: Matter
mesh_terms: D002448:Cell Adhesion; D045744:Cell Line, Tumor; D002465:Cell Movement; D003094:Collagen / Q000737:chemistry; D004338:Drug Combinations; D004847:Epithelial Cells / Q000378:metabolism* / Q000473:pathology; D019573:Epithelium, Corneal / Q000378:metabolism / Q000473:pathology; D005353:Fibronectins / Q000737:chemistry; D005786:Gene Expression Regulation; D020662:Guanine Nucleotide Exchange Factors / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007797:Laminin / Q000737:chemistry; D018994:Myosin Light Chains / Q000235:genetics / Q000378:metabolism*; D024507:Nonmuscle Myosin Type IIA / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D011509:Proteoglycans / Q000737:chemistry; D051057:Proto-Oncogene Proteins c-akt / Q000235:genetics / Q000378:metabolism; D011554:Pseudopodia / Q000378:metabolism* / Q000473:pathology; D064067:Rho Guanine Nucleotide Exchange Factors; D015398:Signal Transduction; D020742:rhoA GTP-Binding Protein / Q000235:genetics / Q000378:metabolism*
chemical_list: C577230:ARHGEF18 protein, human; D004338:Drug Combinations; D005353:Fibronectins; D020662:Guanine Nucleotide Exchange Factors; D007797:Laminin; D018994:Myosin Light Chains; D011509:Proteoglycans; D064067:Rho Guanine Nucleotide Exchange Factors; C059485:matrigel; C066737:RHOA protein, human; D003094:Collagen; D051057:Proto-Oncogene Proteins c-akt; D024507:Nonmuscle Myosin Type IIA; D020742:rhoA GTP-Binding Protein