FARP1, ARHGEF39, and TIAM2 are essential receptor tyrosine kinase effectors for Rac1-dependent cell motility in human lung adenocarcinoma

Cooke, Mariana; Kreider-Letterman, Gabriel; Baker, Martin J.; Zhang, Suli; Sullivan, Neil T.; Eruslanov, Evgeniy; Abba, Martı́n C.; Goicoechea, Silvia M.; Garcı́a-Mata, Rafael; Kazanietz, Marcelo G.

doi:10.1016/j.celrep.2021.109905

Cited by 28 publications

(21 citation statements)

References 79 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To monitor changes in Cdc42 activity in live cells we used a previously characterized dimerization-optimized reporter for activation (DORA) single chain Cdc42 biosensor (generous gift from Yi Wu, UConn Health, Farmington, CT) (Reinhard et al, 2017). Experiments were carried out as previously described for a similar Rac sensor (Baker et al, 2020; Cooke et al, 2021). Briefly, SUM159 cells stably expressing low levels of the FRET-sensor together with a marker for invadopodia (mCherry-cortactin), were treated with PDBu to induce invadopodia.…”

Section: Methodsmentioning

confidence: 99%

ARHGAP17 regulates the spatiotemporal activity of Cdc42 at invadopodia

Kreider-Letterman

Castillo

Mahlandt

et al. 2022

Preprint

Self Cite

View full text Add to dashboard Cite

Cancer cells form actin-rich protrusions called invadopodia that can degrade the extracellular matrix and facilitate tumor invasion and intravasation. Invadopodia formation is regulated by Rho GTPases; however, the molecular mechanisms controlling Rho GTPase signaling at invadopodia are poorly understood. Here, we have identified ARHGAP17, a Cdc42-specific RhoGAP, as a key regulator of invadopodia in breast cancer cells. Using a combination of fixed and live cell imaging, and a Cdc42 sensor optimized for mammalian cells, we have defined a novel ARHGAP17-mediated signaling pathway that controls the spatial and temporal regulation of Cdc42 activity during invadopodia turnover. During assembly, ARHGAP17 localizes to the invadopodia adhesion ring where it restricts the activity of Cdc42 to the invadopodia core. Later, at the start of invadopodia disassembly, ARHGAP17 moves to the core where it inactivates Cdc42 to promote the disassembly of invadopodia in a process that is mediated by its interaction with the Cdc42 effector CIP4. Our results show that ARHGAP17 coordinates when and where Cdc42 is activated during invadopodia assembly and controls the disassembly by terminating the signal.

show abstract

Section: Methodsmentioning

confidence: 99%

ARHGAP17 regulates the spatiotemporal activity of Cdc42 at invadopodia

Kreider-Letterman

Castillo

Mahlandt

et al. 2022

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…In before you begin steps 1–18, and step-by-step method details 1–15, we describe the protocol for analyzing ruffles in live lung adenocarcinoma cells treated with growth factors, such as EGF ( Cooke et al., 2021 ). However, the basic principles can be adapted to other cell types or even other structures, as well as to different treatments.…”

Section: Before You Beginmentioning

confidence: 99%

“…The protocol has applications for dissecting the signaling events linked to ruffling. For complete details on the use and execution of this protocol, please refer to Cooke et al. (2021) .…”

mentioning

confidence: 99%

See 1 more Smart Citation

Quantification of ruffle area and dynamics in live or fixed lung adenocarcinoma cells

Kreider-Letterman

Cooke²,

Goicoechea³

et al. 2022

STAR Protocols

Self Cite

View full text Add to dashboard Cite

“…Here, we describe a method for isolation of EpCAM + cancer cells from fresh surgically resected specimens from lung cancer patients followed by the subsequent identification of specific molecular targets. In our recent study ( Cooke et al, 2021 ), we successfully determined the expression of Rac-GEFs in the purified EpCaM cells using Q-PCR. The current protocol embodies an efficient and valuable tool towards quali-quantitative gene expression analysis in isolated EpCAM + tumor cells and subsequent molecular analysis.…”

Section: Before You Beginmentioning

confidence: 99%