A Rac1-FMNL2 signaling module affects cell-cell contact formation independent of Cdc42 and membrane protrusions.

Publisher: Public Library of Science Country of Publication: United States NLM ID: 101285081 Publication Model: eCollection Cited Medium: Internet ISSN: 1932-6203 (Electronic) Linking ISSN: 19326203 NLM ISO Abbreviation: PLoS One Subsets: MEDLINE

Original Publication: San Francisco, CA : Public Library of Science

Proteins/*metabolism ; Signal Transduction/*physiology ; cdc42 GTP-Binding Protein/*metabolism ; rac1 GTP-Binding Protein/*metabolism; CRISPR-Cas Systems/genetics ; Cell Culture Techniques ; Cell Differentiation ; Cell Line ; Epithelial Cells/cytology ; Epithelial Cells/metabolism ; Formins ; Humans ; Intercellular Junctions/metabolism ; Microscopy, Confocal ; Proteins/antagonists & inhibitors ; Proteins/genetics ; Pseudopodia/physiology ; RNA Interference ; RNA, Small Interfering/metabolism ; cdc42 GTP-Binding Protein/antagonists & inhibitors ; cdc42 GTP-Binding Protein/genetics

De novo formation of epithelial cell-cell contacts relies on actin-based protrusions as well as tightly controlled turnover of junctional actin once cells encounter each other and adhesion complexes assemble. The specific contributions of individual actin regulators on either protrusion formation or junctional actin turnover remain largely unexplored. Based on our previous findings of Formin-like 2 (FMNL2)-mediated control of junctional actin dynamics, we investigated its potential role in membrane protrusions and impact on newly forming epithelial contacts. CRISPR/Cas9-mediated loss of FMNL2 in human MCF10A cells combined with optogenetic control of Rac1 activity confirmed its critical function in the establishment of intercellular contacts. While lamellipodial protrusion rates remained unaffected, FMNL2 knockout cells were characterized by impaired filopodia formation similar to depletion of the Rho GTPase Cdc42. Silencing of Cdc42, however, failed to affect FMNL2-mediated contact formation. Hence, we propose a cell-cell contact-specific and Rac1-mediated function of FMNL2 entirely independent of Cdc42. Consistent with this, direct visualizations of native epithelial junction formation revealed a striking and specifically Rac1- and not Cdc42-dependent recruitment of FMNL2 to newly forming junctions as well as established cell-cell contacts within epithelial sheets.

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0 (FMNL2 protein, human)
0 (Formins)
0 (Proteins)
0 (RNA, Small Interfering)
EC 3.6.5.2 (cdc42 GTP-Binding Protein)
EC 3.6.5.2 (rac1 GTP-Binding Protein)

Date Created: 20180327 Date Completed: 20180706 Latest Revision: 20191210

20240105

PMC5868805

10.1371/journal.pone.0194716

29579104