Lifeact mice for studying F-actin dynamics

Riedl, Julia; Flynn, Kevin C.; Raducanu, Aurelia; Gärtner, Florian; Beck, Gisela; Bösl, Michael R.; Bradke, Frank; Maßberg, Steffen; Aszódi, Attila; Sixt, Michael; Wedlich‐Söldner, Roland

doi:10.1038/nmeth0310-168

Cited by 304 publications

(317 citation statements)

References 3 publications

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“…S6A). We used Lifeact (which is known to bind both G-actin and F-actin biochemically) for targeting PLEKHG3 to F-actin because Lifeact is used as a marker to visualize F-actin in living cells and is known not to interfere with the actin dynamics in cells (22,23). The illuminated regions showed the induction of lamellipodia, and the direction of the cells eventually changed because of the accumulation of PLEKHG3 (Fig.…”

Section: Plekhg3 Localizes To the Leading Edge And Controls Cell Migrmentioning

confidence: 99%

PLEKHG3 enhances polarized cell migration by activating actin filaments at the cell front

Nguyen

Park

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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Cells migrate by directing Ras-related C3 botulinum toxin substrate 1 (Rac1) and cell division control protein 42 (Cdc42) activities and by polymerizing actin toward the leading edge of the cell. Previous studies have proposed that this polarization process requires a local positive feedback in the leading edge involving Rac small GTPase and actin polymerization with PI3K likely playing a coordinating role. Here, we show that the pleckstrin homology and RhoGEF domain containing G3 (PLEKHG3) is a PI3K-regulated Rho guanine nucleotide exchange factor (RhoGEF) for Rac1 and Cdc42 that selectively binds to newly polymerized actin at the leading edge of migrating fibroblasts. Optogenetic inactivation of PLEKHG3 showed that PLEKHG3 is indispensable both for inducing and for maintaining cell polarity. By selectively binding to newly polymerized actin, PLEKHG3 promotes local Rac1/Cdc42 activation to induce more local actin polymerization, which in turn promotes the recruitment of more PLEKHG3 to induce and maintain cell front. Thus, autocatalytic reinforcement of PLEKHG3 localization to the leading edge of the cell provides a molecular basis for the proposed positive feedback loop that is required for cell polarization and directed migration.PLEKHG3 | cell polarity | F-actin binding | positive feedback | PI3K

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Section: Plekhg3 Localizes To the Leading Edge And Controls Cell Migrmentioning

confidence: 99%

PLEKHG3 enhances polarized cell migration by activating actin filaments at the cell front

Nguyen

Park

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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“…We used the following mouse lines: Srf floxed (SRF fl/fl ) (Parlakian et al, 2004); PDGFb-iCreER (Claxton et al, 2008); the Cre reporter Rosa26-mTmG (mTmG) (Muzumdar et al, 2007); and LifeAct-GFP (Riedl et al, 2010). Mice were genotyped by PCR.…”

Section: Micementioning

confidence: 99%

“…The LifeAct-GFP mouse expresses an actin-binding protein fused to eGFP that labels filamentous actin specifically (Riedl et al, 2010). Importantly, the LifeAct-GFP transgene is strongly expressed by the retinal endothelium, allowing visualization of the actin cytoskeleton specifically in the vasculature (Fraccaroli et al, 2012).…”

Section: Endothelial Actin Filament Organization Defects In Srfmentioning

confidence: 99%

SRF selectively controls tip cell invasive behavior in angiogenesis

et al. 2013

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SUMMARYEfficient angiogenic sprouting is essential for embryonic, postnatal and tumor development. Serum response factor (SRF) is known to be important for embryonic vascular development. Here, we studied the effect of inducible endothelial-specific deletion of Srf in postnatal and adult mice. We find that endothelial SRF activity is vital for postnatal growth and survival, and is equally required for developmental and pathological angiogenesis, including during tumor growth. Our results demonstrate that SRF is selectively required for endothelial filopodia formation and cell contractility during sprouting angiogenesis, but seems dispensable for vascular remodeling. At the molecular level, we observe that vascular endothelial growth factor A induces nuclear accumulation of myocardinrelated transcription factors (MRTFs) and regulates MRTF/SRF-dependent target genes including Myl9, which is important for endothelial cell migration in vitro. We conclude that SRF has a unique function in regulating migratory tip cell behavior during sprouting angiogenesis. We hypothesize that targeting the SRF pathway could provide an opportunity to selectively target tip cell filopodia-driven angiogenesis to restrict tumor growth.

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“…However, one of the drawbacks of our transfection system is that the expression of the protein of interest is limited to one or two cells per acinus, limiting the possibility to study the behavior of the actin cytoskeleton in groups of cell. Recently, a transgenic mouse expressing GFP-life act has been generated (Riedl et al, 2010). These mice are superior to the GFP-actin transgenic mice in terms of viability, level of protein expression and cellular toxicity and represent a formidable tool to study several aspect of the involvement of actin in various cellular processes.…”

Section: Actin Cytoskeletonmentioning

confidence: 99%