Modulation of lamellipodial structure and dynamics by NO-dependent phosphorylation of VASP Ser239

Lindsay, Susan L.; Ramsey, Sara; Aitchison, Michael; Renné, Thomas; Evans, Thomas J.

doi:10.1242/jcs.003061

Cited by 54 publications

(62 citation statements)

References 42 publications

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“…affects the interaction of VASP with actin; however, the crucial phosphorylation sites and kinases involved are still unclear (Barzik et al, 2005;Harbeck et al, 2000;Laurent et al, 1999;Lindsay et al, 2007). VASP phosphorylation also modulates other proteinprotein interactions.…”

Section: Introductionmentioning

confidence: 99%

“…Additionally, VASP phosphorylation status appears to modulate subcellular protein distribution. It has been postulated that VASP phosphorylation by PKA controls protein targeting to the cell-cell junctions of adherent cells (Benz et al, 2008;Comerford et al, 2002), but others have shown that PKG activity might control VASP localization (Lindsay et al, 2007;Smolenski et al, 2000).…”

Section: Introductionmentioning

confidence: 99%

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Differential VASP phosphorylation controls remodeling of the actin cytoskeleton

Benz

Blume

Seifert

et al. 2009

Journal of Cell Science

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156

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Proteins of the Enabled/vasodilator-stimulated phosphoprotein (Ena/VASP) family link signal transduction pathways to actin cytoskeleton dynamics. VASP is substrate of cAMP-dependent, cGMP-dependent and AMP-activated protein kinases that primarily phosphorylate the sites S157, S239 and T278, respectively. Here, we systematically analyzed functions of VASP phosphorylation patterns for actin assembly and subcellular targeting in vivo and compared the phosphorylation effects of Ena/VASP family members. Methods used were the reconstitution of VASP-null cells with `locked' phosphomimetic VASP mutants, actin polymerization of VASP mutants in vitro and in living cells, site-specific kinase-mediated VASP phosphorylation, and analysis of the endogenous protein with phosphorylation-status-specific antibodies. Phosphorylation at S157 influenced VASP localization, but had a minor impact on F-actin assembly. Phosphorylation of the S157-equivalent site in the Ena/VASP family members Mena and EVL had no effect on the ratio of cellular F-actin to G-actin. By contrast, VASP phosphorylation at S239 (and the equivalent site in Mena) or T278 impaired VASP-driven actin filament formation. The data show that VASP functions are precisely regulated by differential phosphorylation and provide new insights into cytoskeletal control by serine/threonine kinase-dependent signaling pathways.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Differential VASP phosphorylation controls remodeling of the actin cytoskeleton

Benz

Blume

Seifert

et al. 2009

Journal of Cell Science

Self Cite

156

234

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“…Ser153 are defective in restriction of S. flexneri spread Activity of VASP is regulated in part by phosphorylation on serines and threonines (Benz et al, 2009;Blume et al, 2007;Döppler et al, 2013;Lindsay et al, 2007). Phosphorylation is known to occur on four residues, corresponding to Ser153, Ser235, Thr274 and Ser322 of the murine protein.…”

Section: Vasp Derivatives Defective In Phosphorylation Atmentioning

confidence: 99%

Vasodilator-stimulated phosphoprotein restricts cell-to-cell spread of Shigella flexneri at the cell periphery

2015

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Shigella spp. are intracellular bacterial pathogens that cause diarrhoeal disease in humans. Shigella utilize the host actin cytoskeleton to enter cells, move through the cytoplasm of cells and pass into adjacent cells. Ena/VASP family proteins are highly conserved proteins that participate in actin-dependent dynamic cellular processes. We tested whether Ena/VASP family members VASP (vasodilator-stimulated phosphoprotein), Mena (mammalian-enabled) or EVL (Ena-VASP-like) contribute to Shigella flexneri spread through cell monolayers. VASP and EVL restricted cell-to-cell spread without significantly altering actin-based motility, whereas Mena had no effect on these processes. Phosphorylation of VASP on Ser153, Ser235 and Thr274 regulated its subcellular distribution and function. VASP derivatives that lack the Ena/VASP homology 1 (EVH1) domain or contain a phosphoablative mutation of Ser153 were defective in restricting S. flexneri spread, indicating that the EVH1 domain and phosphorylation on Ser153 are required for this process. The EVH1 domain and Ser153 of VASP were required for VASP localization to focal adhesions, and localization of VASP to focal adhesions and/or the leading edge was required for restriction of spread. The contribution of the EVH1 domain was from both the donor and the recipient cell, whereas the contribution of Ser153 phosphorylation was only from the donor cell. Thus, unlike host proteins characterized in Shigella pathogenesis that promote bacterial spread, VASP and EVL function to limit it. The ability of VASP and EVL to limit spread highlights the critical role of focal adhesion complexes and/or the leading edge in bacterial passage between cells.

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“…Importantly, Ser239 within the EVH2 VASP domain is a preferred phosphorylation site for PKG, functioning as a biological marker for cGMP signalling in intestinal (Deguchi et al, 2002) and other cells (Krause et al, 2003;Yaroslavskiy et al, 2005). Cyclic GMP-dependent VASP phosphorylation inhibits membrane protrusion formation in normal cells (Krause et al, 2003;Lindsay et al, 2007). Accordingly, in colorectal cancer cells VASP Ser239 phosphorylation induced by ligand activation of GCC signalling through cGMP and PKG induces rapid disassembly (less than 10 minutes) of invasive and migratory membrane organelles (Zuzga et al, 2011).…”

Section: Control Of Invasive Cell Shapementioning

confidence: 99%