A novel mode of capping protein-regulation by twinfilin

Johnston, Adam B.; Hilton, Denise M; McConnell, Patrick I.; Johnson, Britney; Harris, Meghan T; Simone, Avital; Amarasinghe, Gaya K.; Cooper, John A.; Goode, Bruce L.

doi:10.7554/elife.41313

Cited by 39 publications

(58 citation statements)

References 61 publications

(127 reference statements)

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“…Indeed, our results reveal substantial differences among the CPI motifs with respect to V-1 and F-actin interactions. Our results are consistent with previous studies with a more limited scope (1,21). These biochemical differences among the CPI-motif peptides may reflect differences in their cellular functions, which could be tested in the future.…”

Section: Discussionsupporting

confidence: 92%

“…We therefore investigated the kinetics of V-1 dissociation from the CP:V-1 complex induced by different CPI-motif-protein peptides. In previous reports, the CPI-motif regions of CARMIL1 and twinfilin were found to increase the dissociation rate of V-1 from CP (7,8,21).…”

Section: Effect Of Cpi-motif Peptides On Dissociation Of V-1 From Cpmentioning

confidence: 80%

“…Myotrophin (V-1, P58546, Homo sapiens) was expressed and purified as described (20). N-carboxytetramethylrhodamine (TAMRA)-V-1 (C45S, C83S) was prepared as described (21). Rabbit skeletal muscle alpha-actin (actin, P68135, Oryctolagus cuniculus) was prepared as described (22).…”

Section: Proteins and Peptidesmentioning

confidence: 99%

“…The peptides were derived as follows, with peptide boundaries and UniProtKB designations for the proteins: Capping protein Arp2/3 Myosin-I Linker (CARMIL1 G969-A1005, Q5VZK9, Homo sapiens), Capping protein, Arp2/3 and Myosin-I linker protein 3 (CARMIL3 E959-M994, Q8ND23, Homo sapiens), WASH complex subunit 2C (WASHCAP A990-R1026, Q9Y4E1, Homo sapiens), CapZ-interacting protein (CapZIP V140-R176, Q6JBY9, Homo sapiens), CD2-associated protein (CD2AP D473-H509, Q9Y5K6, Homo sapiens), SH3 domain-containing kinase-binding protein 1 (CIN85 L463-S499, Q96B97, Homo sapiens), Casein kinase 2interacting protein-1 (Pleckstrin homology domain-containing family O member 1, CKIP-1/PLEKHO R137-M173, Q53GL0, Homo sapiens), and Twinfilin-1 (Twf-1 H317-D350, Q91YR1, Mus musculus). The Twf-1 peptide was identical to one previously described (21). The CPI-motif peptide from human CARMIL2 was tested, but it had poor solution properties and was not included in this study.…”

Section: Proteins and Peptidesmentioning

confidence: 99%

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Comparative analysis of CPI-motif regulation of biochemical functions of actin capping protein

McConnell¹,

Mekel²,

Козлов³

et al. 2020

Preprint

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The heterodimeric actin capping protein (CP) is regulated by a set of proteins that contain CP-interacting (CPI) motifs. Outside of the CPI motif, the sequences of these proteins are unrelated and distinct. The CPI motif and surrounding sequences are conserved within a given protein family, when compared to those of other CPI-motif protein families. Using biochemical assays with purified proteins, we compared the ability of CPI-motif-containing peptides from different protein families to a) bind to CP, b) allosterically inhibit barbed-end capping by CP, and c) allosterically inhibit interaction of CP with V-1, another regulator of CP.We found large differences in potency among the different CPI-motif-containing peptides, and the different functional assays showed different orders of potency. These biochemical differences among the CPI-motif peptides presumably reflect interactions between CP and CPI-motif peptides involving amino-acid residues that are conserved but are not part of the strictly defined

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Section: Discussionsupporting

confidence: 92%

Section: Effect Of Cpi-motif Peptides On Dissociation Of V-1 From Cpmentioning

confidence: 80%

Section: Proteins and Peptidesmentioning

confidence: 99%

Section: Proteins and Peptidesmentioning

confidence: 99%

See 2 more Smart Citations

Comparative analysis of CPI-motif regulation of biochemical functions of actin capping protein

McConnell¹,

Mekel²,

Козлов³

et al. 2020

Preprint

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“…Therefore, we focused on its other interaction partners. Twinfilin binds CP through its C-terminal tail 41,42,58 , and both twinfilin and CP localize to lamellipodia in mammalian cells 16,43,59,60 . Whereas CP localizes to the distal edge of lamellipodia 16 , the precise localization pattern of twinfilin has not been reported.…”

Section: Twinfilin Controls the Dynamics Of Cp In Cellsmentioning

confidence: 99%

Twinfilin uncaps filament barbed ends to promote turnover of lamellipodial actin networks

Hakala

Wioland

Tolonen

et al. 2019

Preprint

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Coordinated polymerization of actin filaments provides force for cell migration, morphogenesis, and endocytosis. Capping Protein (CP) is central regulator of actin dynamics in all eukaryotes. It binds actin filament (F-actin) barbed ends with high affinity and slow dissociation kinetics to prevent filament polymerization and depolymerization. In cells, however, CP displays remarkably rapid dynamics within F-actin networks, but the underlying mechanism has remained enigmatic. We report that a conserved cytoskeletal regulator, twinfilin, is responsible for CP's rapid dynamics and specific localization in cells. Depletion of twinfilin led to stable association of CP with cellular F-actin arrays and its treadmilling throughout leading-edge lamellipodium. These were accompanied by diminished F-actin disassembly rates. In vitro single filament imaging approaches revealed that twinfilin directly promotes dissociation of CP from filament barbed ends, while allowing subsequent filament depolymerization. These results uncover an evolutionary conserved bipartite mechanism that controls how actin cytoskeleton-mediated forces are generated in cells.Coordinated polymerization of actin filaments (F-actin) generates pushing force, which drives the protrusion of lamellipodia at the leading edge of migrating cells, and contributes to the formation of plasma membrane invaginations in endocytic processes 1-4 . A large array of actin-binding proteins regulates the dynamics and organization of actin filaments in these processes, but only few (<10) of them are conserved in evolution from protozoan parasites to animals 5 . Among these 'core' actin-binding proteins are the heterodimeric Capping Protein (CP) and twinfilin.Generation of membrane protrusions requires that a subset of actin filament barbed ends is capped to funnel the assemblycompetent actin monomers to a limited number of growing barbed ends 6-8 . CP is the most prominent actin filament barbed end capping protein in most organisms and cell types 9,10 . It is an essential component of in vitro reconstituted actin-based motility system 11 and actin-based processes in cells [12][13][14] . Moreover, its inhibition in animal non-muscle cells disturbs actin-based lamellipodial morphology, protrusion and cell migration [15][16][17][18] . In addition, CP plays an important role in controlling the length and the density of branches within actin filament networks nucleated by the Arp2/3 complex 19 .The activities of CP are controlled by several proteins 9,10 . V-1/ myotrophin binds and sequesters CP with nanomolar affinity and inhibits its capping activity 20-23 . Moreover, the capping protein interaction (CPI)-motif containing proteins, such as CARMILs (capping protein, Arp2/3 and myosin-I linker protein) 24 , interact with CP to reduce its affinity to both actin filament barbed ends [25][26][27] and to V-1 22 through an allosteric mechanism 25,28 . Depletion of CARMILs and other CPI motif containing proteins disrupts the subcellular localization of CP 29-33 . Thus, it was suggested...

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Cytoskeleton | Actin-Capping and Severing Proteins

Maiti

2021

Encyclopedia of Biological Chemistry III

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A novel mode of capping protein-regulation by twinfilin

Cited by 39 publications

References 61 publications

Comparative analysis of CPI-motif regulation of biochemical functions of actin capping protein

Comparative analysis of CPI-motif regulation of biochemical functions of actin capping protein

Twinfilin uncaps filament barbed ends to promote turnover of lamellipodial actin networks

Cytoskeleton | Actin-Capping and Severing Proteins

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