Actin-binding domains mediate the distinct distribution of two Dictyostelium Talins through different affinities to specific subsets of actin filaments during directed cell migration

Tsujioka, Masatsune; Uyeda, Taro Q.P.; Iwadate, Yoshiaki; Patel, Hitesh; Shibata, Keitaro; Yumoto, Tenji; Yonemura, Shigenobu

doi:10.1371/journal.pone.0214736

Cited by 3 publications

(3 citation statements)

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“…The third term is a representation of the adhesion energy between the membrane and the cortex. Proteins connecting the membrane to the cortex have been found around the boundary of migrating cells [ 18 , 47 ]. Their contribution to adhesion energy is commonly modeled using linear elastic spring with spring constant κ [ 17 , 18 , 23 , 24 ].…”

Section: Methodsmentioning

confidence: 99%

A role for myosin II clusters and membrane energy in cortex rupture for Dictyostelium discoideum

et al. 2022

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Blebs, pressure driven protrusions of the cell membrane, facilitate the movement of eukaryotic cells such as the soil amoeba Dictyostelium discoideum, white blood cells and cancer cells. Blebs initiate when the cell membrane separates from the underlying cortex. A local rupture of the cortex, has been suggested as a mechanism by which blebs are initiated. However, much clarity is still needed about how cells inherently regulate rupture of the cortex in locations where blebs are expected to form. In this work, we examine the role of membrane energy and the motor protein myosin II (myosin) in facilitating the cell driven rupture of the cortex. We perform under-agarose chemotaxis experiments, using Dictyostelium discoideum cells, to visualize the dynamics of myosin and calculate changes in membrane energy in the blebbing region. To facilitate a rapid detection of blebs and analysis of the energy and myosin distribution at the cell front, we introduce an autonomous bleb detection algorithm that takes in discrete cell boundaries and returns the coordinate location of blebs with its shape characteristics. We are able to identify by microscopy naturally occurring gaps in the cortex prior to membrane detachment at sites of bleb nucleation. These gaps form at positions calculated to have high membrane energy, and are associated with areas of myosin enrichment. Myosin is also shown to accumulate in the cortex prior to bleb initiation and just before the complete disassembly of the cortex. Together our findings provide direct spatial and temporal evidence to support cortex rupture as an intrinsic bleb initiation mechanism and suggests that myosin clusters are associated with regions of high membrane energy where its contractile activity leads to a rupture of the cortex at points of maximal energy.

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Section: Methodsmentioning

confidence: 99%

A role for myosin II clusters and membrane energy in cortex rupture for Dictyostelium discoideum

et al. 2022

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“…In Dictyostelium , TalA is implicated in cell-substrate adhesion and cytokinesis in the unicellular vegetative state, while TalB is involved in cell motility, force transmission, and development during the transition to a multicellular state ( Niewohner et al, 1997 ; Tsujioka et al, 2004 ; Tsujioka et al, 2008 ). Furthermore, both molecules have also been shown to bind to actin and possess conserved FERM and I/LWEQ domains similar to their Metazoan counterparts ( Tsujioka et al, 2008 ; Tsujioka et al, 2019 ). Intriguingly, double knockouts of talA and talB in Dictyostelium demonstrated a severe defect in cell-substrate adhesion and migration ( Tsujioka et al, 2008 ), suggesting a critical role for both TalA and TalB in both cellular processes.…”

Section: Cell-substrate Adhesions In Organisms Evolutionarily Distant...mentioning

confidence: 99%

An evolutionary and physiological perspective on cell-substrate adhesion machinery for cell migration

Morales

Xue

Roh-Johnson

2022

Front. Cell Dev. Biol.

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Cell-substrate adhesion is a critical aspect of many forms of cell migration. Cell adhesion to an extracellular matrix (ECM) generates traction forces necessary for efficient migration. One of the most well-studied structures cells use to adhere to the ECM is focal adhesions, which are composed of a multilayered protein complex physically linking the ECM to the intracellular actin cytoskeleton. Much of our understanding of focal adhesions, however, is primarily derived from in vitro studies in Metazoan systems. Though these studies provide a valuable foundation to the cell-substrate adhesion field, the evolution of cell-substrate adhesion machinery across evolutionary space and the role of focal adhesions in vivo are largely understudied within the field. Furthering investigation in these areas is necessary to bolster our understanding of the role cell-substrate adhesion machinery across Eukaryotes plays during cell migration in physiological contexts such as cancer and pathogenesis. In this review, we review studies of cell-substrate adhesion machinery in organisms evolutionary distant from Metazoa and cover the current understanding and ongoing work on how focal adhesions function in single and collective cell migration in an in vivo environment, with an emphasis on work that directly visualizes cell-substrate adhesions. Finally, we discuss nuances that ought to be considered moving forward and the importance of future investigation in these emerging fields for application in other fields pertinent to adhesion-based processes.

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“…More recently it was shown that talin A binds actin filaments pre-stretched by myosin II especially at the uropod of developed cells, while talin B is enriched at the anterior cell pole excluded from those regions [195].…”

Section: Talb0mentioning

confidence: 99%

Mechanics and Dynamics of <i>Dictyostelium discoideum</i> adhesion

Kamprad¹

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The amoeba Dictyostelium discoideum (D.d.) is an excellent laboratory model system for studying amoeboid adhesion. In its natural habitat, D.d. lives in the soil and hunts for bacteria, an environment which is very heterogeneous where surface properties changes rapidly. This phenomenon requires a high degree of flexibility from the adhesion apparatus of D.d.. To block amoeboid adhesion of D.d., first studies used polyethylene glycol (PEG) "brushes" as surface functionalization. In the present study, it was shown that gels of PEG are a good alternative, and it was possible to identify that developmental stage of D.d., axenic background and cytoskeletal fluorescence labelling can reduce the effect of surface passivation.In a second set of experiments conducted in the present work, the focus lay on the adhesion versatility of D.d. to a variety of substrates, which bases on several non-specific driving forces. To specify these forces, model substrates made of silicon were used, which influence DLVO forces based in their layered structure. It was found that Van der Waals forces, hydrophobic effects and electrostatic interactions are involved in D.d. adhesion. Besides membrane-and glycocalyx-based interactions, these forces are complemented by additional structures, individual adhesion bonds and adhesion bond clusters of the transmembrane protein SadA (Substrate Adhesion-deficient A), while D.d. does not possess integrins and thus focal contacts.Furthermore, actin foci, dynamic structures of freshly polymerised actin in the ventral membrane, have been shown to be involved in amoeboid adhesion and migration. In the present work, it was possible to analyse SadA adhesion bonds and actin foci both from a mechanical point of view employing step spectroscopy based on single cell force spectroscopy, but also in a more dynamical approach using reflection interference contrast and total internal reflection microscopy. Several established axenic laboratory strains as well as corresponding mutants deficient for components of actin-adhesomeand endocytosis-signalling were analysed.The following two phenomena could were observed: I) reduction of the adhesion forces and step as well as spot density for inhibition of the Arp2/3 complex II) manipulation of adhesion apparatus (anchoring protein TalA, transmembrane protein SadA) increases the number of actin foci, with a simultaneous reduction in adhesion force.Finally a quantification of the role of actin foci in adhesion and endocytosis was achieved. It was possible to show that actin foci exist independently of clathrin-mediated endocytosis, which also affects the lifetime of actin foci. Furthermore, clathrin-containing structures could also be detected after the appearance of actin foci, which may stabilise adhesive structures.It was shown that D.d. is a very versatile organism that adapts substrate-dependently to the environment by non-specific adhesion. Furthermore, it could also be shown that specific adhesion of D.d. works very similarly to that of integrins. v Chapter 0 -Conten...

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Actin-binding domains mediate the distinct distribution of two Dictyostelium Talins through different affinities to specific subsets of actin filaments during directed cell migration

Cited by 3 publications

References 60 publications

A role for myosin II clusters and membrane energy in cortex rupture for Dictyostelium discoideum

A role for myosin II clusters and membrane energy in cortex rupture for Dictyostelium discoideum

An evolutionary and physiological perspective on cell-substrate adhesion machinery for cell migration

Mechanics and Dynamics of <i>Dictyostelium discoideum</i> adhesion

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