An attenuating role of a WASP-related protein, WASP-B, in the regulation of F-actin polymerization and pseudopod formation via the regulation of RacC during Dictyostelium chemotaxis

Chung, Chang Y.; Feoktistov, Alexander; Hollingsworth, Ryan J.; Rivero, Francisco; Mandel, Nicole S.

doi:10.1016/j.bbrc.2013.06.022

Cited by 8 publications

(12 citation statements)

References 23 publications

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“…In order to obtain spatial and temporal information about the activity of small GTPases in Dictyostelium, fluorescent probes based on GTP-binding domains (GBDs) from effector molecules were introduced. For example, GBDs from Dictyostelium WASP and PakB fused with fluorescent proteins have been used to monitor the dynamics of RacC and Rac1, respectively (Han et al 2006;Chung et al 2013;Veltman et al 2012), but the specificity of these probes is possibly broader (Rivero and Xiong 2016). Within the Ras family, Ras-binding domain (RBD) of mammalian Raf1 or RalGDS fused to GFP was used as biosensors for activated RasG (Kae et al 2004).…”

Section: Discussionmentioning

confidence: 99%

Quantitative imaging of Rac1 activity in Dictyostelium cells with a fluorescently labelled GTPase-binding domain from DPAKa kinase

Marinović

Šoštar

Filić

et al. 2016

Histochem Cell Biol

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Small Rho GTPases are major regulators of the actin cytoskeleton dynamics in eukaryotic cells. Sophisticated tools used to investigate their activity in living cells include probes based on fluorescence resonance energy transfer (FRET), bimolecular fluorescence complementation, and photoactivation. However, such methods are of limited use in quickly migrating cells due to a short time available for image acquisition leading to a low signal-to-noise ratio. Attempts to remedy this effect by increasing the intensity of illumination are restricted by photobleaching of probes and the cell photosensitivity. Here we present design and characterization of a new fluorescent probe that selectively binds to active form of Rac1 GTPases, and demonstrate its superior properties for imaging in highly motile Dictyostelium cells. The probe is based on the GTPase-binding domain (GBD) from DPAKa kinase and was selected on the basis of yeast two-hybrid screen, GST pull-down assay and FRET measurements by fluorescence lifetime imaging microscopy. DPAKa(GBD) probe binds specifically to GTP-bound Rac1 at the cell membrane and features a low cytoplasmic background. The main advantage of DPAKa(GBD) in comparison with similar probes is its finely graded intensity distribution along the entire plasma membrane, which enables quantitative measurements of the Rac1 activity in different parts of the membrane. Finally, expression of DPAKa(GBD) induces no adverse effects on cell growth, motility and cytokinesis.

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Section: Discussionmentioning

confidence: 99%

Quantitative imaging of Rac1 activity in Dictyostelium cells with a fluorescently labelled GTPase-binding domain from DPAKa kinase

Marinović

Šoštar

Filić

et al. 2016

Histochem Cell Biol

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“…Although the SCAR/WAVE complex appears to play a predominant role in directing cell migration in Dictyostelium , WASP can complement its loss to regulate pseudopod formation [291]. In addition, a WASP-related protein, WASP-B, has been shown to negativity regulate RacC activity that controls pseudopod extension during chemotaxis to cAMP [296]. …”

Section: Chemotactic Network Of Dictyostelium and Leukocytesmentioning

confidence: 99%

Moving towards a paradigm: common mechanisms of chemotactic signaling in Dictyostelium and mammalian leukocytes

Artemenko

Lampert

Devreotes

2014

Cell. Mol. Life Sci.

181

221

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Chemotaxis, or directed migration of cells along a chemical gradient, is a highly coordinated process that involves gradient sensing, motility, and polarity. Most of our understanding of chemotaxis comes from studies of cells undergoing amoeboid-type migration, in particular the social amoeba Dictyostelium discoideum and leukocytes. In these amoeboid cells the molecular events leading to directed migration can be conceptually divided into four interacting networks: receptor/G protein, signal transduction, cytoskeleton, and polarity. The signal transduction network occupies a central position in this scheme as it receives direct input from the receptor/G protein network, as well as feedback from the cytoskeletal and polarity networks. Multiple overlapping modules within the signal transduction network transmit the signals to the actin cytoskeleton network leading to biased pseudopod protrusion in the direction of the gradient. The overall architecture of the networks, as well as the individual signaling modules are remarkably conserved between Dictyostelium and mammalian leukocytes, and the similarities and differences between the two systems are the subject of this review.

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“…We encounter a similar situation in Dictyostelium ( Figure 2 ), where it is also not possible to draw a sharp dividing line between the pathways driven by individual GTPases, neither in interaction nor in functional assays. The yeast-two-hybrid (Y2H) and pull-down assays that use CRIB motifs from different effectors as baits are especially prone to show interactions with multiple GTPases, and vice versa [ 59 , 80 , 81 , 82 , 83 , 84 , 85 ]. Refining the existing map of functional interactions between individual GTPases and their effectors in Dictyostelium therefore remains an important task for future research.…”

Section: Comparative Analysis Of the Rho Signalling In Dictyostelium And Mammalian Cellsmentioning

confidence: 99%

Regulation of the Actin Cytoskeleton via Rho GTPase Signalling in Dictyostelium and Mammalian Cells: A Parallel Slalom

Filić

Mijanović

Putar

et al. 2021

Cells

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Both Dictyostelium amoebae and mammalian cells are endowed with an elaborate actin cytoskeleton that enables them to perform a multitude of tasks essential for survival. Although these organisms diverged more than a billion years ago, their cells share the capability of chemotactic migration, large-scale endocytosis, binary division effected by actomyosin contraction, and various types of adhesions to other cells and to the extracellular environment. The composition and dynamics of the transient actin-based structures that are engaged in these processes are also astonishingly similar in these evolutionary distant organisms. The question arises whether this remarkable resemblance in the cellular motility hardware is accompanied by a similar correspondence in matching software, the signalling networks that govern the assembly of the actin cytoskeleton. Small GTPases from the Rho family play pivotal roles in the control of the actin cytoskeleton dynamics. Indicatively, Dictyostelium matches mammals in the number of these proteins. We give an overview of the Rho signalling pathways that regulate the actin dynamics in Dictyostelium and compare them with similar signalling networks in mammals. We also provide a phylogeny of Rho GTPases in Amoebozoa, which shows a variability of the Rho inventories across different clades found also in Metazoa.

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An attenuating role of a WASP-related protein, WASP-B, in the regulation of F-actin polymerization and pseudopod formation via the regulation of RacC during Dictyostelium chemotaxis

Cited by 8 publications

References 23 publications

Quantitative imaging of Rac1 activity in Dictyostelium cells with a fluorescently labelled GTPase-binding domain from DPAKa kinase

Quantitative imaging of Rac1 activity in Dictyostelium cells with a fluorescently labelled GTPase-binding domain from DPAKa kinase

Moving towards a paradigm: common mechanisms of chemotactic signaling in Dictyostelium and mammalian leukocytes

Regulation of the Actin Cytoskeleton via Rho GTPase Signalling in Dictyostelium and Mammalian Cells: A Parallel Slalom

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